Cephem compounds and pharmaceutical compositions containing the same

ABSTRACT

A cephem compound, wherein the cephem ring has a substituent at the 3-position, which substituent is shown by the formula II, wherein Het is a mono- or polycyclic heterocyclic group comprising one or more hetero atoms selected from the group consisting of N, O and S which may be the same or different from each other; R 1  is hydrogen, an optionally substituted lower alkyl or an optionally substituted lower alkenyl; A is an optionally substituted lower alkylene, an optionally substituted lower alkenylene or a single bond; B is an optionally substituted imino or a single bond; or A and B taken together may form a single bond; and D is a single bond or a group of the formula (a):                    
     The cephem compounds of the present invention are useful as antibiotic agents.

This application has been filed under 35 U.S.C. § 371 fromPCT/JP97/01161, filed Apr. 4, 1997.

TECHNICAL FIELD

The present invention relates to novel cephem compounds, a process forpreparing the same, intermediates therefor, and pharmaceuticalcompositions containing the compounds.

BACKGROUND ART

Compounds having an optionally substituted pyridiniomethyl group at the3-position of the cephem ring have been disclosed in patent applicationssuch as Japanese Patent Publication (KOKAI) 60-237090 (WO 8505106, EP160969A2), Japanese Patent Publication (KOKOKU) 1-44190 and alsoJapanese Patent Publication (KOKOKU) 6-70068 (EP 64740B1, U.S. Pat. No.5,071,979), Japanese Patent Publication (KOKOKU) 2-44476 (EP 159011B1,U.S. Pat. No. 4,833,242), etc. However, there have not been reportedcompounds wherein a pyridinium ring is substituted with a heterocyclicgroup having a substituent of the formula —CONHCN or its analogues.

Although a huge number of antibiotics have been marketed so far, thedevelopment and characterization of compounds with higher antibioticactivity have been continuously demanded so as to cope with theappearance of multiple drug resistant bacteria and to provide for thediversification of therapy forms. In particular, it has been demanded todevelop cephem compounds of broad spectrum which show a long bloodhalf-life and have an excellent in vivo dynamics such as transfer to atissue.

DISCLOSURE OF INVENTION

The present inventors have intensively studied with a purpose fordeveloping novel cephem compounds with superior characteristics andfound that cephem compounds wherein the cephem ring has apyridiniomethyl group at the 3-position, and wherein the pyridinium ringis substituted with a heterocyclic group having a substituent -CONHCN oran analogue thereof have an excellent in vivo dynamics properties.

Thus, the present invention provides a cephem compound wherein thecephem ring has a substituent at the 3-position, which substituent isshown by the formula II:

wherein

Het is a mono- or polycyclic heterocyclic group comprising one or morehetero atoms selected from the group consisting of N, O and S which maybe the same or different from each other; R¹ is hydrogen, an optionallysubstituted lower alkyl or an optionally substituted lower alkenyl; A isan optionally substituted lower alkylene, an optionally substitutedlower alkenylene or a single bond; B is an optionally substituted iminoor a single bond; and D is a single bond or a group of the formula:

or a salt or a hydrate thereof. The above-mentioned cephem compounds,salts or hydrates may be hereinafter referred to as the compound of thepresent invention.

The compound of the present invention is preferably represented by theformula I:

wherein Acyl is an acyl, and Het, R¹, A, B and D are as defined above,or an ester, a salt, or a hydrate thereof.

Acyl in the formula I is preferably a group of the formula III:

wherein X is CH or N; Y is an optionally protected amino; and Z is anoptionally substituted hydrocarbon group.

Het in the formula I or II is preferably a 5- or 6-membered trivalenthetero cyclic group comprising one to four hetero atoms selected fromthe group consisting of N, O and S which may be the same or differentfrom each other, and, more preferably, a pyrrolyl group of the formulaIV:

Further, A in the formula I or II is preferably a single bond or a vinylgroup; B is a single bond; and D is a single bond.

Example of preferable compounds of the formula I include those whereinAcyl is a group of the formula III:

(wherein X is CH or N, Y is an optionally protected amino and Z ishydrogen or an optionally substituted hydrocarbon group); Het is a 5- or6-membered hetero cyclic group comprising one to four hetero atomsselected from the group consisting of N, O and S which may be the sameor different from each other; A is a single bond or a vinyl group; B isa single bond; and D is a single bond, or an ester, a salt, or a hydratethereof.

Terms herein used are defined below.

Throughout the present specification, the term “cephem compound” refersto a class of compounds having a double bond between the 3- and4-positions of the cepham ring and named according to the nomenclatureshown under the heading “cephem” in The Journal of the American ChemicalSociety, 84, 3400 (1962). The present invention encompasses compounds ofthe formula I pharmaceutically acceptable esters, salts, or hydratesthereof (i.e., esters of Compound I, salts of Compound I, salts of anester of Compound I, or hydrates thereof). The signal “⁻” in —COO⁻ atthe 4-position of a compound of the formula I indicates that acarboxylate anion forms an intramolecular salt by making a pair with thepyridinium cation on the substituent at the 3-position. When thecarboxyl group is not ionized, the pyridinium cation can form a saltwith an anion or a counter ion on a side-chain. The present inventionencompasses all of these embodiments. The “S” at the 1-position of thecephem ring may be oxidized.

The term “mono- or polycyclic heterocyclic group” in the definition of“Het” includes the both aromatic and non-aromatic mono- or polycyclicheterocyclic groups, which is bound to the adjacent three groups. In thecase of a monocyclic heterocyclic group, examples of a preferredaromatic heterocyclic group include 5- to 6-membered cyclic groups suchas furan, thiophene, tetrazole, pyrrole, pyrazole, imidazole, oxazole,thiazole, pyridine, oxazine and triazine. Examples of a preferrednon-aromatic heterocyclic group include 5- to 7-membered groups such aspyrrolidine, thiazolidine, oxazolidine, imidazolidine, thiazoline,oxazoline, imidazoline, piperidine, piperazine, morpholine,thiomorpholine, oxadiazoline, and dioxane. Among them, a monocyclicheterocyclic group comprising one or two hetero atoms selected from Nand S is more preferable, and pyrrole is most preferred.

Preferred examples of polycyclic heterocyclic groups include thosewherein benzene ring, pyridine ring, pyrazine ring, pyridazine ring,pyrimidine ring, or the like, is condensed to an above-mentionedmonocyclic aromatic heterocyclic group, such as benzothiophene, indole,benzothiazole, benzofuran, and benzimidazole. Those wherein Het is boundto the 4-position of pyridinium ring are preferred.

The term “lower alkyl” in the definition of “R¹” refers to a straight orbranched C₁₋₆alkyl groups, such as methyl, ethyl, propyl, isopropyl,butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl,tert-pentyl, 1-ethylpropyl, hexyl, isohexyl, 1,1-dimethylbutyl,2,2-dimethylbutyl, 3,3-dimethylbutyl, 2-ethylbutyl, and the like.C₁₋₄alkyl group such as methyl, ethyl, propyl, isopropyl, butyl,isobutyl, and the like, are preferred. The lower alkyl group may besubstituted by a substituent(s) selected from, for example, loweralkenyl group (e.g., C₂₋₆alkenyl group such as vinyl, butenyl, propenyl,etc.); cycloalkyl group (e.g., C₃₋₇cycloalkyl group such as cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, etc.); aryl group(e.g., C₆₋₁₀aryl group such as phenyl, naphthyl, etc., which aryl groupmay be further substituted by hydroxy, C₁₋₄alkyl such as methyl orethyl, or C₁₋₄alkoxy such as methoxy or ethoxy); aromatic heterocyclicgroup (e.g., 5- or 6-membered aromatic heterocyclic group comprising 1to 4 hetero atoms selected from N, O, S, and the like, such as furyl,thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl,imidazolyl, pyrazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl,1,3,4-oxadiazolyl, furazanyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl,1,3,4- thiadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, tetrazolyl,pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl; bi- ortricyclic aromatic condensed heterocyclic group comprising 1 to 5 heteroatoms selected from N, O, S, and the like, which is formed by condensingone or two 5- or 6-membered aromatic heterocyclic groups comprising 1 to4 hetero atoms selected from N, O, S, and the like or one or two benzenerings, such as benzofuranyl, isobenzofuranyl, benzo[b]thienyl, indolyl,isoindolyl, 1H-inzdazolyl, bonzimidazolyl, benzoxazolyl,1,2-benzisoxazolyl, benzothiazolyl, 1 ,2-benzisothiazolyl,1H-benzotriazolyl, quinolyl, isoquinolyl, cinnolinyl, quinazolinyl,quinoxalinyl, phthaladinyl, naphthylidinyl, purinyl, putelidinyl,carbazolyl, α-carbolinyl, β-carbolinyl, γ-carbolinyl, acridinyl,phenoxazinyl, phenothiazinyl, phenazinyl, phenoxathiinyl, thianthrenyl,phenathridinyl, phenathrolinyl, indolydinyl, pyrrolo[1,2-b]pyridazinyl,pyrazolo[1,5-a]pyridyl, imidazo[1,2-a]pyridyl, imidazo[1,5-a]pyridyl,imidazo[1,2-b]pyridazinyl, imidazo[1,2-a]pyrimidinyl,1,2,4-triazolo[4,3-a]pyridyl, 1,2,4-triazolo[4,3-b]pyridazinyl);non-aromatic heterocyclic group (e.g., 4- or 6-membered non-aromaticheterocyclic group comprising 1 to 3 hetero atoms selected from N, O, S,and the like, such as oxiranyl, azetidinyl, oxetanyl, thiethanyl,pyrrolidinyl, tetrahydrofuryl, thiolanyl, piperidyl, tetrahydropyranyl,morpholinyl, thiomorpholinyl, piperadinyl, and the like); amino group;mono- or di-lower alkyl amino group (e.g., mono- or diC₁₋₆alkylaminogroup such as methylamino, ethylamino, dimethylamino, and the like);tri-lower alkylammonium group (e.g., triC₁₋₆alkylammonium group such astrimethylammonium, triethylammonium, tripropylammonium, and the like);amidino group; acyl group (e.g., C₁₋₆alkanoyl group such as formyl,acetyl, propionyl, and the like); carbamoyl group; mono- or di-loweralkylcarbamoyl group (e.g., mono- or diC₁₋₆alkylcarbamoyl group such asmethylcarbamoyl, ethylcarbamoyl, dimethylcarbamoyl, and the like);sulfamoyl group; mono- or di-lower alkyl sulfamoyl group (e.g., mono- ordiC₁₋₆alkylsulfamoyl group such as methylsulfamoyl, ethylsulfamoyl,dimethylsulfamoyl, and the like); carboxyl group; lower alkoxycarbonylgroup (e.g., C₁₋₆alkoxycarbonyl group such as methoxycarbonyl,ethoxycarbonyl, and the like); hydroxyl group; lower alkoxy group (e.g.,C₁₋₆alkoxy group such as methoxy, ethoxy, and the like); loweralkenyloxy group (e.g., C₂₋₆alkenyloxy group such as allyloxy,2-buthenyloxy, and the like); cycloalkyloxy group (e.g.,C₃₋₇cycloalkyloxy group such as cyclopropyloxy, cyclobutyloxy,cyclopentyloxy, cyclohexyloxy, cycloheptyloxy, and the like); aralkyloxygroup (e.g., C₇₋₁₀aralkyloxy group such as benzyloxy, phenethyloxy, andthe like); aryloxy group (e.g., C₆₋₁₀aryloxy group such as phenoxy,naphthyloxy, and the like); mercapto group; lower alkylthio group (e.g.,C₁₋₆alkylthio group such as methylthio, ethylthio, and the like);aralkylthio group (e.g., C₇₋₁₀aralkylthio group such as benzylthio,phenethylthio, and the like); arylthio group (e.g., C₆₋₁₀arylthio groupsuch as phenylthio, naphthylthio, and the like); sulfo group; cyanogroup; azide group; nitro group; nitroso group; halogen (e.g., fluorine,chlorine, iodine, and the like). The number of substituent is preferably1 to 3 and when there are more than one substituents, they may be thesame or different from each other.

The term “lower alkenyl” refers to a straight or branched C₂₋₆alkenylgroup such as allyl, propenyl, butenyl, pentenyl, and the like, andallyl is preferred. The lower alkenyl group may be substituted by asubstituent(s) similar to those mentioned above for lower alkyl group.

The term “lower-alkylene” in the definition of “A” refers to a groupderived from the above-mentioned lower alkyl groups, for example,methylene, ethylene, butylene, propylene, pentylene, and the like, andmethylene and ethylene are preferred. The lower alkylene group can besubstituted by a substituent(s) similar to those mentioned above forlower alkyl group.

The term “lower alkenylene” refers to a group derived from theabove-mentioned lower alkenyl groups, for example, vinylene, butenylene,propenylene, and the like, and vinylene is preferred. The loweralkenylene group can be substituted by a substituent(s) similar to thosementioned above for lower alkyl group.

The “acyl group” represented by Acyl refers to an acyl group known as asubstituent for the 6-amino group of penicillin derivatives as well asthe 7-amino group of cephem compounds. Examples of such acyl groupsinclude those derived from organic carboxylic acids such as formylgroup; alkylcarbonyl group (alkanoyl group), preferably,(C₁-C₆)alkyl-carbonyl group (e.g., acetyl, propionyl, butyryl,isobutyryl, valeryl, isovaleryl, pivaloyl, hexanoyl, and the like);(C₃-C₅)alkenoyl group (e.g., acryloyl, chrotonoyl, maleoyl, and thelike); (C₃-C₁₀)cycloalkyl-carbonyl group (e.g., cyclopropylcarbonyl,cyclobutylcarbonyl, cyclopentylcarbonyl, cyclohexylcarbonyl,cycloheptylcarbonyl, adamantylcarbonyl, and the like);(C₅-C₆)cycloalkenyl-carbonyl group (e.g., cyclopentenylcarbonyl,cyclopentadienylcarbonyl, cyclohexenylcarbonyl, cyclohexadienylcarbonyl,and the like); arylcarbonyl group (aroyl group), preferably,(C₆-C₁₄)aryl-carbonyl group (e.g., benzoyl, 1- or 2-naphthoyl, and thelike); aralkyl carbonyl group, preferably, (C₇-C₁₉)aralkyl-carbonylgroup (e.g., phenylacetyl, phenylpropionyl, α,α,α-triphenylacetyl,2-phenetylcarbonyl, 1- or 2-naphthylmethylcarbonyl, benzhydrylcarbonyl,and the like); 5- or 6-membered aromatic heterocyclic carbonyl group(e.g., 2- or 3-thenoyl, 2- or 3-furoyl, nicotinoyl, isonicotinoyl, 4- or5-thiazolylcarbonyl, 1,2,4-thiadiazol-3- or1,2,4-thiadiazol-5-yl-carbonyl, and the like); 5- or 6-membered aromaticheterocyclic acetyl group (e.g., 2- or 3-thienylacetyl, 2- or3-furylacetyl, 4-thiazolylacetyl, 1,2,4-thiadiazol-3-yl-acetyl,1-tetrazolylacetyl, and the like); alkoxycarbonyl group, preferably,(C₁-C₆)alkoxy-carbonyl group (e.g., methoxycarbonyl, ethoxycarbonyl,propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl,tert-butoxycarbonyl, and the like); aryloxycarbonyl group, preferably,(C₆-C₁₄)aryloxy-carbonyl group (e.g., phenoxycarbonyl, 1- or2-naphtoxycarbonyl, and the like); aralkyloxycarbonyl group, preferably,(C₇-C₁₉)aralkyloxycarbonyl group (e.g;, benzyloxycarborryl, and thelike); aminoalkylcarbonyl group (e.g., amino C₁₋₆alkyl-carbonyl groupsuch as glycyl, aranyl, valyl, leucyl, isoleucyl, seryl, threonyl,cysteinyl, cystynyl, methionyl, asparaginyl, glutamyl, lysyl, arginyl,phenylglycyl, phenylalanyl, tyrosyl, histidyl, tryptophanyl, prolyl,2-aminoethylcarbonyl, 3-aminopropylcarbonyl, and the like);monoalkylaminoalkylcarbonyl group (e.g.,monoC₁₋₆alkylamino-C₁₋₆alkyl-carbonyl group such asmethylaminomethylcarbonyl, 2-ethylaminoethylcarbonyl, and the like); anddialkylaminoalkylcarbonyl group (e.g.,diC₁₋₆alkylamino-C₁₋₆alkyl-carbonyl group such asdimethylaminomethylcarbonyl, diethylaminomethylcarbonyl, and the like).

These acyl group may be substituted by one to three substituentsselected from amino, nitro, halogen (e.g., fluorine, chlorine, bromine,and the like), hydroxy, oxo, carbamoyl group, (C₁-C₄)alkyl group (e.g.,methyl, ethyl, propyl, isopropyl, butyl, and the like), (C₁-C₄)alkoxygroup (e.g., methoxy, ethoxy, propoxy, butoxy, and the like), optionallyesterified carboxyl group (e.g., (C₁-C₆)alkoxycarbonyl group such asmethoxycarbonyl, ethoxycarbonyl, and the like), (C₁-C₄)alkoxyimino groupwhich is optionally substituted by carboxyl or halogen (e.g.,methoxyimino, ethoxyimino, carboxymethoxyimino,1-carboxy-1-mehylethoxyimino, fluoromethoxyimino, fluoroethoxyimino, andthe like), hydroxyimino group, and4-ethyl-2,3-dioxopiperadinocarbonylamino group.

The heterocyclic group in the 5- or 6-membered aromatic heterocycliccarbonyl group and 5- or 6-membered aromatic heterocyclic acetyl groupas defined above refers to an aromatic heterocyclic group comprising oneto four hetero atoms selected from the group consisting of optionallyoxidized nitrogen atom, oxygen atom, optionally mono- or dioxidizedsulfur atom, and the like, and examples other than those set forth aboveinclude pyrrole, imidazole, pyrazole, pyrimidine, pyrazine, pyridazine,indole, isothiazole, oxazole, isoxazole, and triazole.

Preferred examples of Acyl include those shown by the formula (III)wherein X is CH or N; Y is an optionally protected amino; and Z ishydrogen or an optionally substituted hydrocarbon group.

Examples of amino-protecting groups in the definition of Y include anappropriate group used in the field of β-lactam- and peptide chemistry.Preferred amino-protecting group includes formyl, chloroacetyl,tert-butoxycarbonyl, benzyloxycarbonyl, p-methoxybenzyloxycarbonyl,p-nitrobenzyloxycarbonyl, 2-trimethylsilylethoxycarbonyl,2,2,2-trichloroethoxycarbonyl, and trityl.

Examples of a hydrocarbon group in the definition of “Z” include loweralkyl, lower alkenyl, lower alkynyl, cycloalkyl, aralkyl, di- ortriaryl-methyl and aryl. The lower alkyl group is a straight or branchedalkyl group of, preferably, 1 to 6 carbon atoms, such as methyl, ethyl,n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl,n-hexyl, and the like. The lower alkenyl group is a straight or branchedalkenyl group of, preferably, 2 to 6 carbon atoms, such as allyl,propenyl, butenyl, pentenyl, and the like. The lower alkynyl group is astraight or branched alkynyl group of, preferably, 2 to 6 carbon atoms,such as propynyl, butynyl, pentynyl, and the like. The cycloalkyl groupis preferably a cycloalkyl group of 3 to 6 carbon atoms, such ascyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like. Thearalkyl group is preferably a group of 7 to 10 carbon atoms, such asbenzyl, and the like. The di- or triaryl-methyl group is preferably adi- or tri(C₆₋₁₀aryl)-methyl group, such as benzhydryl,di(p-tolyl)methyl, trityl, tri(p-tolyl)methyl, and the like. The arylgroup is a group of 6 to 10 carbon atoms, such as phenyl, and the like.

The hydrocarbon group shown by “Z” may be substituted by one to threesubstituents selected from, for example, carboxyl group;C₁₋₆alkoxy-carbonyl group such as methoxycarbonyl, ethoxycarbonyl, andthe like; carbamoyl group; C₁₋₆alkylthio group such as methylthio,ethylthio, and the like; sulfamoyl group; amino group; hydroxy group;cyano group; carbamoyloxy group; and halogen such as fluorine, chlorine,and the like. Examples of preferred Z include hydrogen, (C₁-C₃)loweralkyl group and a lower alkyl group substituted by one or 2 substituentsselected from halogen and carboxyl group (e.g., fluoromethyl,fluoroethyl, carboxypropyl, etc.)

Ester derivatives of a compound or an intermediate of the presentinvention are those formed through the esterification of a carboxylgroup(s) in the molecule, and are usable as a synthetic intermediate ora non-toxic metabolic ester which is apt to undergo hydrolysis in vivo.

Examples of ester derivatives usable as a synthetic intermediate includeoptionally substituted C₁₋₆alkyl ester, C₂₋₆alkenyl ester,C₃₋₁₀cycloalkyl ester, C₃₋₁₀cycloalkyl-C₁₋₆alkyl ester, optionallysubstituted C₆₋₁₀aryl ester, optionally substituted C₇₋₁₂aralkyl ester,diC₆₋₁₀aryl-methyl ester, triC₆₋₁₀aryl-methyl ester, substituted silylester, and the like.

Examples of metabolic ester residues include acetoxymethyl group,1-acetoxyethyl group, 1-acetoxypropyl group; pivaloyloxymethyl group,1-isopropyloxycarbonyloxyethyl group, 1-cyclohexyloxycarbonyloxyethylgroup, phthalidyl group, (2-oxo-5-methyl-1,3-dioxol-4-yl)methyl, and thelike.

When the —COO⁻ group at the 4-position of Compound I is esterified, theester residue can be, for example, a group of the formula VIII:

wherein R⁷ is hydrogen, an alkyl group, a cycloalkyl group or acycloalkylalkyl group; R⁸ is hydrogen, an alkyl group, a cycloalkylgroup, an alkoxy group, a cycloalkyloxy group, a cycloalkylalkyl group,an alkenyloxy group, or a phenyl group; a phthalidyl group;(2-oxo-5-methyl-1,3-dioxol-4-yl)methyl group; an alkoxyalkyl group; analkylthioalkyl group; a tert-butyl group; a 2,2,2-trichloroethyl group;a benzyl group; a p-methoxybenzyl group; a p-nitrobenzyl group; abenzhydryl group; a trityl group; a trimethylsilyl group; or an allylgroup.

In the above definition, the alkyl group or the alkyl moiety incycloalkylalkyl group, alkoxyalkyl group and alkylthioalkyl group canbe, for example, a straight or branched group of 1 to 6 carbon atoms(e.g., methyl, ethyl, propyl, isopropyl, butyl, 2,2-dimethylpropyl,etc.), and the cycloalkyl group or the cycloalkyl moiety incycloalkyloxy group or cycloalkylalkyl group can be, for example, acycloalkyl group of 3 to 7 carbon atoms (e.g., cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl, etc). Examples of alkoxy group oralkoxy moiety in alkoxyalkyl group include a straight or a branchedchain alkoxy group of 1 to 10 carbon atoms (e.g., methoxy, ethoxy,propoxy, isopropoxy, butoxy, hexyloxy, decyloxy, etc.) Examples ofalkenyloxy group include a straight or a branched chain alkenyloxy groupof 2 to 7 carbon atoms (e.g., allyloxy, etc.)

As a salt of the compound of the present invention, pharmaceuticallyacceptable salts are preferred, such as those formed with an inorganicbase, an organic base, an inorganic acid, an organic acid, a basic oracidic amino acid, and intra-molecular salts. Examples of preferredsalts formed with an inorganic base include alkali metal salts such assodium or potassium salts; alkaline-earth metal salts such as calcium ormagnesium salts; alminium salts; and ammonium salts. Examples ofpreferred salts formed with an organic base include those formed withtrimethylamine, triethylamine, pyridine, picoline, ethanolamine,diethanolamine, triethanolamine, dicyclohexylamine,N,N′-dibenzylethylenediamine, procaine, 2-phenylethylbenzylamine,trishydroxymethylaminomethane, polyhydroxyalkylamine,N-methylglucosamine, and the like. Examples of preferred salts formedwith an inorganic acid include those formed with hydrochloric acid,hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, and thelike. Examples of preferred salts formed with an organic acid includethose formed with formic acid, acetic acid, trifluoroacetic acid,fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid,succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid,p-toluenesulfonic acid, and the like. Examples of preferred salts formedwith basic amino acid include those formed with arginine, lysine,ornithine, histidine, and the like. Examples of preferred salts formedwith an acidic amino acid include those formed with aspartic acid,glutamic acid, and the like.

Among the salts, the base-addition salts (i.e., salts with inorganic ororganic base, or basic amino acid) are those which can be formed at the4-carboxyl group of the cephem ring or an acidic group such as carboxylgroup, sulfo group, hydroxyl group, or the like, on the side chain, ifany. The acid-addition salts (i.e., salts with inorganic or organicacid, or acidic amino acid) means those which can be formed at a basicgroup such as amino group, monoalkylamino group, dialkylamino group,cycloalkylamino group, arylamino group, aralkylamino group, N-containingheterocyclic group, or the like, of a compound of the present invention,if any. Acid addition salts also include those having a counter ion suchas chloride ion, bromide ion, sulfate ion, p-toluenesulfonate ion,methanesulfonate ion, trifluoroacetate ion, or the like, which areformed when an organic or inorganic acid (1 mole) is attached to thesite of a compound of the present invention where an intramolecular saltis formed between the 4-carboxylate moiety (COO⁻) and the pyridiniumcation on the 3-side chain.

The hydrate of the present invention refers to a mono- or dihydrate.They are obtainable by selecting an appropriate drying method.

The compound of the present invention can be prepared according to aknown method in the field of β-lactam. The typical processes areprovided below.

[Production Method 1]

A compound of the formula I, or an ester or a salt thereof can beprepared by reacting a cephem compound of the formula V:

wherein R⁴ is a carboxy-protecting group, R⁵ is a hydroxy group, anacyloxy group, a carbamoyloxy group, a substituted carbamoyloxy group,or a halogen atom, or a salt thereof with a pyridine derivative of theformula VI:

wherein R¹, A, B, D and Het are as defined above, or a salt thereof, andoptionally deprotecting the reaction product.

In the reaction, Compound V or its salt (hereinafter, they may bereferred to as Compound V) and a pyridine derivative VI or its salt(hereinafter, they may be referred to as Compound VI) are reacted togive Compound I through the nucleophilic substitution reaction. CompoundV can be easily obtained in accordance with a known method such as thosedescribed in Japanese Patent Publication (KOKAI) 231684/1985 or JapanesePatent Publication (KOKAI) 149682/1987, or a method equivalent thereto.Compound VI can be prepared in a manner shown in the working examplesbelow.

The nucleophilic substitution of Compound V by Compound VI is normallycarried out in a solvent. Solvents useable in the reaction are ethers(dioxane, tetrahydrofuran, diethylether, etc.), esters (ethyl formate,ethyl acetate, n-butyl acetate, etc.), halogenated hydrocarbons(dichloromethane, chloroform, carbon tetrachloride, 1,2-dichloroethane,etc.), hydrocarbons (n-hexane, benzene, toluene, etc.), amides(formamide, N,N-dimethylformamide, etc.), ketones (acetone, methyl ethylketone, etc.), nitrites (acetonitrile, propionitrile, etc.), and alsodimethyl sulfoxide, sulfolane, hexamethylphosphoramide, and water, whichare used alone or in combination as a mixed solvent. Further, alcoholssuch as methanol, ethanol, n-propanol, isopropanol, ethylene glycol,2-methoxyethanol, can be used.

When Compound VI is liquid, it can be used in a large excess (e.g., 10to 200-fold moles) to Compound V so that it can serve as a solvent. Insuch a case, Compound VI can be used in combination with any one or moresolvents above to give a mixed solvent.

When R⁵ in Compound V is an acyloxy group, a carbamoyloxy group or asubstituted carbamoyloxy group, more preferred solvent is water, or amixed solvent of water and a water-miscible organic solvent. Preferredexamples of the organic solvent include acetone, methyl ethyl ketone,acetonitrile, and the like. The amount of Compound VI is normallybetween about 1 to 5 moles, preferably about 1 to 3 moles, based on 1mole of Compound V. The reaction is conducted at temperature range ofabout 10 to 100° C., preferably about 30 to 80° C. The reaction timedepends on the kinds of Compound V, compound VI, or the solvent,reaction temperature, or the like, but is normally from about tensminutes to several hours, preferably from about 1 to 5 hours. Thereaction is advantageously conducted at the pH range of 2 to 8,preferably about neutral, i.e. pH 5 to 8. This reaction easily proceedsin the presence of 2 to 30 equivalents of iodides or thiocyanates.Examples of such salts include sodium iodide, potassium iodide, sodiumthiocyanate, potassium thiocyanate, and the like. The reaction can beallowed to proceed smoothly by adding quaternary ammonium salts having asurface activity action such as trimethylbenzylammonium bromide,triethylbenzylammonium bromide, triethylbenzylammonium hydroxide, andthe like, in addition to the above salts.

When R⁵ in Compound V is a hydroxyl group, the reaction can be effectedin the presence of an organophosphorous compound according to the methoddescribed, for example, in Japanese Patent Publication (KOKAI) 58-43979(corresponding to U.S. Pat. Nos. 4,642,365 and 4,801,703).

Preferred solvents usable in the reaction include, for example, theabove-mentioned ethers, esters, halogenated hydrocarbons, hydrocarbons,amides, ketones, nitriles and sulfoxides, which are used alone or incombination. Particularly, dichloromethane, acetonitrile,dimethylformamide, dimethyl sulfoxide, a mixed solvent ofdimethylformamide and acetonitrile, and a mixed solvent ofdichloromethane and acetonitrile would lead to good results. The amountof Compound VI or a salt thereof, and that of the organophosphorouscompound is preferably from about 1 to 5 moles and about 1 to 10 moles,more preferably from about 1 to 3 moles and about 1 to 6 moles,respectively, based on 1 mole of Compound V. The reaction is conductedat temperature range of about −80 to 50° C., preferably about −40 to 40°C. The reaction time is normally from about 30 minutes to 48 hours,preferably from about 1 to 24 hours. An organic base can be added in thereaction system. Examples of the organic base include amines such astriethylamine, tri(n-butyl)amine, di(n-butyl)amine, diisobutylamine,dicyclohexylamine, and the like. The amount of the base is preferablyabout 1 to 5 moles based on 1 mole of Compound V.

When R⁵ in compound V is a halogen atom (preferably iodine), preferablesolvents are the above ethers, esters, halogenated hydrocarbons,hydrocarbons, amides, ketones, nitrites, alcohols, water, sulfoxides,and the like. The amount of Compound VI is normally from about 1 to 5moles, preferably from about 1 to 3 moles, based on 1 mole of CompoundV. The reaction is conducted at temperature range of about 0 to 80° C.,preferably about 20 to 60° C. The reaction time is normally from about30 minutes to 15 hours, preferably from about 1 to 5 hours. The reactioncan be facilitated in the presence of a dehydrohalogenating agent.Examples of dehydrohalogenating agent usable in the reaction includedeacidifying agents such as inorganic bases (e.g. sodium carbonate,potassium carbonate, calcium carbonate, sodium hydrogencarbonate, etc.),tertiary-amines (e.g. triethylamine, tri(n-propyl)amine,tri(n-butyl)amine, diisopropylethylamine, cyclohexyldimethylamine,pyridine, lutidine, etc.) and alkylene oxides (e.g. propylene oxide,epichlorohydrin, etc.), but Compound VI itself can be used as thedehydrohalogenating agent. In this case, Compound VI is used in theamount of 2 moles or more based on 1 mole of Compound V.

[Production Method 2]

A compound wherein Acyl in the formula I is shown by the formula III canalso be produced through the etherification by reacting a hydroxyiminoderivative of the formula VII:

wherein the respective symbols are as defined above, an ester, or a saltthereof with a compound of the formula ZOH (wherein Z is as definedabove) or a reactive derivative thereof. The reactive derivatives of ZOHare those capable of replacing a hydrogen atom of the hydroxyiminocompound VII with Z and include, for example, a compound of the formulaZR⁶ (wherein R⁶ is a leaving group such as a halogen atom, amono-substituted sulfonyloxy group, etc.). Examples of themono-substituted sulfonyloxy group include C₁₋₆alkylsulfonyloxy groupand C₆₋₁₀arylsulfonyloxy group, such as methanesulfonyloxy,ethanesulfonyloxy, benzenesulfonyloxy, p-toluenesulfonyloxy, and thelike.

The hydroxyimino compound VII can be synthesized by the method describedherein or those known in the art.

The compound ZOH and reactive derivatives thereof can be easilysynthesized by a known method, for example, those described in JapanesePatent Publication (KOKAI) Nos. 60-231684 and 62-149682) or analoguesthereof.

When using ZOH, the hydroxyimino compound VII is reacted with a compoundZOH by using an appropriate dehydrating agent to synthesize Compound I.Examples of the dehydrating agent used for this purpose includephoshorous oxychloride, thionyl chloride, dialkyl azodicarboxylate(normally used in combination with phosphine),N,N′-dicyclohexylcarbodiimide, and the like. Preferred dehydrating agentis diethyl azocarboxylate in combination with triphenylphosphine. Thereaction using diethyl azocarboxylate in combination withtriphenylphosphine is normally conducted in an anhydrous solvent. Forexample, the above-mentioned ethers and hydrocarbons are used. Thecompound ZOH, ethyl azodicarboxylate and triphenylphosphine are used inthe amount of about 1 to 1.5 moles based on 1 mole of the hydroxyiminocompound VII. The reaction takes about several tens minutes to a fewhours at temperature range of about 0 to 50° C.

When using ZR⁶, the reaction between ZR⁶ and the hydroxyimino compoundVII is a normal etherification reaction which is conducted in a solvent.As the solvent, there can be used the above-mentioned solvents such asethers, esters, halogenated hydrocarbons, hydrocarbons, amides, ketones,nitrites, alcohols, water, or the like, or a mixed solvent. The solventis preferably a mixed solvent of water and a water-miscible solvent, forexample, water-containing methanol, water-containing ethanol,water-containing acetone, water-containing dimethyl sulfoxide, or thelike. The reaction is also allowed to proceed smoothly in the presenceof an appropriate base. Examples of the base include inorganic base suchas alkaline metal salts including sodium carbonate, sodium bicarbonate,potassium carbonate, etc., and alkaline metal hydroxides includingsodium hydroxide, potassium hydroxide, etc. This reaction can also beconducted in a buffer (e.g. phosphate buffer) at pH 7.5 to 8.5. Thecompound ZR⁶ and the base are used at about 1 to 5 moles and about 1 to10 moles, preferably about 1 to 3 moles and about 1 to 5 moles,respectively, on the basis of 1 mole of compound VII. The reactiontemperature can be in the range of about −30 to 100° C., preferablyabout 0 to 80° C. The reaction time is about 10 minutes to 15 hours,preferably about 30 minutes to 5 hours.

A function group(s) such as amino, hydroxy, carboxy, or the like, can beprotected with an appropriate protecting group when effecting theaforementioned respective reaction.

The method of deprotection and purification for producing the compoundof the present invention will be hereinafter explained.

Deprotection Method:

For example, a monohalogenoacetyl group (e.g. chloroacetyl, bromoacetyl)can be removed by using thiourea; an alkoxycarbonyl group (e.g.methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl) can be removed byusing an acid (e.g. hydrochloric acid); an aralkyloxycarbonyl group(e.g. benzyloxycarbonyl, p-methylbenzyloxycarbonyl,p-nitrobenzyloxycarbonyl) can be removed by catalytic reduction;2,2,2-trichloroethoxycarbonyl can be removed by using zinc and an acid(e.g. acetic acid); 2-methylsulfonylethyl ester can be removed by usingan alkali; an aralkyl ester (e.g. benzyl ester, p-methoxybenzyl ester,p-nitrobenzyl ester) can be removed by using an acid (e.g. formic acid,trifluoroacetic acid, AlCl₃, TiCl₄) or by catalytic reduction; a2,2,2-trichloroethyl ester can be removed by using zinc and an acid(e.g. acetic acid); and a silyl ester (e.g. trimethylsilyl ester,tert-butyldimethylsilyl ester) can be removed by using water alone.

Purification Method:

The compound of the present invention or a synthetic intermediatethereof obtained by the above-mentioned or other production methods canbe isolated and purified according to known methods includingextraction, column chromatography, precipitation, recrystallization, andthe like. Further, the isolated compound can then be converted intodesired physiologically acceptable salts by a known method.

The compound of the present invention is useful as a drug, especially, avaluable antibiotic because it shows an antibacterial activity of broadspectrum, a long blood half-life, and an excellent in vivo dynamics.Therefore, the compound can be used directly or indirectly for thepurpose of preventing or treating various diseases caused by pathogenicmicroorganisms in human and mammals (e.g. mouse, rat, rabbit, canine,cat, bovine, swine), for example, sinopulmonary infection and urinaryinfection. The antibacterial spectra are characteristic in the followingpoints.

(1) It is highly active on various Gram-negative bacteria.

(2) It is highly active on Gram-positive bacteria.

(3) It is highly active on methicillia resistant Staphylococcus aurous(MRSA).

(4) It is highly active on Pseudomonas which is insensitive to thetreatment with a normal cephalosporin antibiotic.

(5) It is also highly active on various Gram-negative bacteria capableof producing β-lactamase (e.g. genus Escherichia, genus Enterobacter,genus Serratia, genus Proteus, etc.).

Microorganisms of the genus Pseudomonas have so far been treated withaminoglycoside antibiotics such as amikacin, gentamicin, and the like.The compound of the present invention has a great advantage over theaminoglycosides because the former exerts antibacterial activitiesequivalent to the latter with by far the less toxicity to human andanimals.

The compound of the present invention can be orally or parenterallyadministered in the form of solid preparations (e.g. tablets, capsules,granules, powders, etc.) or liquid preparations (e.g. syrups,injections, etc.) in association with pharmaceutically acceptablecarriers.

As the pharmaceutically acceptable carriers, there can be used variousorganic or inorganic carriers which have been commonly used as materialsfor pharmaceutical preparations. In case of the solid preparation,excipients, lubricants, binders and disintegrators, and in case of theliquid preparation, solvents, solubilizers, suspending agents,isotonicities, buffering agents and soothing agents, can beappropriately combined. If necessary, preparation additives such asantiseptics, antioxidants, colorants and sweetening agents can also beused according to conventional methods. Preferred examples of theexcipient include lactose, sucrose, D-mannitol, starch, crystallinecellulose, light anhydrous silicic acid, and the like. Preferredexamples of the lubricant include magnesium stearate, calcium stearate,talc, colloidal silica, and the like. Preferred examples of the binderinclude crystalline cellulose, sucrose, D-mannitol, dextrin,hydroxypropyl cellulose, hydroxypropylmethyl cellulose,polyvinylpyrrolidone, and the like. Preferred examples of thedisintegrator include starch, carboxymethyl cellulose, calciumcarboxymethyl cellulose, sodium cross carboxymethyl cellulose, sodiumcarboxymethyl starch, and the like. Preferred examples of the solventinclude distilled water for injection, alcohol, propylene glycol,macrogol, sesame oil, corn oil, and the like. Preferred examples of thesolubilizer include polyethylene glycol, propylene glycol, D-mannitol,benzyl benzoate, ethanol, tris-aminomethane, cholesterol,triethanolamine, sodium carbonate, sodium citrate, and the like.Preferred examples of the suspending agent include surfactants such asstearyltriethanolamine, sodium lauryl sulfate, laurylaminopropionicacid, lecithin, benzalkonium chloride, benzetonium chloride, glycerinmonostearate, and the like; and hydrophilic polymer such as polyvinylalcohol, polyvinylpyrrolidone, sodium carboxymethyl cellulose, methylcellulose, hydroxymethyl cellulose, hydroxyethyl cellulose,hydroxypropyl cellulose, and the like. Preferred examples of theisotonicity include sodium chloride, glycerin, D-mannitol, and the like.Preferred examples of the buffering agent include buffer solutions ofphosphate, acetate, carbonate and citrate. Preferred examples of thesoothing agent include benzyl alcohol, and the like. Preferred examplesof the antiseptic include paraoxybenzoates, chlorobutanol, benzylalcohol, phenethyl alcohol, dehydroacetic acid, sorbic acid, and thelike. Preferred examples of the antioxidant include sulfite, ascorbate,and the like. It is also possible to obtain a preparation having anantibacterial activity of broader spectrum by mixing other activeingredient(s) (e.g. β-lactam antibiotic).

The compound of the present invention can be used for preventing andtreating bacterial infections such as respiratory infection, urinaryinfection, pyogenic disease, biliary infection, intestinal infection,obstetric infection, otolaryngologic infection and surgical infection ofhuman and other mammals. Although the dosage varies depending on theconditions and weight of patients and administration method, the dailydose of an active ingredient for adult for parenteral administration cangenerally be about 0.5-80 mg/kg, preferably about 2-40 mg/kg, which isadministered in one to three divisions, by intravenous or intramuscularinjection. For oral administration, the daily dose of an activeingredient can be 1-100 mg/kg, preferably about 2.5-50 mg/kg, which isadministered in one to three divisions.

BEST MODE FOR CARRYING OUT THE INVENTION

The abbreviations used in the following Preparations and Examples are asfollows:

THF: tetrahydrofuran, DBU: 1,8-diazabicyclo[5,4,0]undecene, DMF:dimethylformamide, DMSO: dimethyl sulfoxide, DIBAH: diisobutylaluminumhydride, TMS: trimethylsilyl, Me: methyl, Et: ethyl, iPr: isopropyl,^(t)Bu: tert-butyl, Ph: phenyl, MsCl: methanesulfonyl chloride.

(Protecting Group)

Boc: tert-butyloxycarbonyl

Im: imidazolyl

BH: diphenylmethyl

PMB: p-methoxybenzyl

POM: tert-butylcarbonyloxymethyl

With respect to the expression of the compounds, the figure underlinedordinarily corresponds to a compound of the same number in the chemicalformula, wherein the figure is underlined by “˜”.

(1) To a solution of 1 (7.74 ml, 70 mmol) and ethyl formate (11.3 ml,0.14 moles) in 150 ml of benzene was added MeONa (powder) (7.6 g, 0.14moles), and the mixture was stirred at room temperature for 1.5 hours.The mixture was refluxed for additional 30 minutes and benzene wasdistilled away under reduced pressure. The resulting residue wasdissolved in 150 ml of THF. To the solution were added acetic acid (8.6ml, 0.15 moles) and diethyl aminomalonate (14.8 g, 70 mmol) in series,and the mixture was stirred at room temperature for 2 hours. Thereaction solution was poured into NaHCO₃-ice-cold water and extractedwith ethyl acetate. The extract was washed with water, dried andconcentrated under reduced pressure to obtain crude 3 as residue. Theresidue was purified by silica gel chromatography (CH₂Cl₂/ethylacetate=3/1) to obtain 14.0 g of 3 (yield 65.3%).

Compound 3:

NMR(CDCl₃) δ: 1.33(6H,t,J=7.0 Hz), 4.32(4H,q,J=7.0 Hz), 4.65(1H,d,J=8.4Hz), 5.86(1H,d,J=7.8 Hz), 6.99˜7.10(1H,m), 7.10(2H,d,J=5.6 Hz),8.73(2H,d,J=5.6 Hz)

(2) To 3 (21.56 g, 70.4 mmol) was added 80 g of polyphosphoric acid, andthe mixture was heated on an oil bath at 90° C. for 1 hour. The reactionsolution was cooled, added to ice-cold water and neutralized withNa₂CO₃. The insoluble materials were collected by filtration, dissolvedin ethyl acetate, dried over anhydrous magnesium sulfate andconcentrated under reduced pressure. The resulting crystalline residuewas washed with ethyl acetate to obtain 6.02 g of 4 (yield 39.5%) as acrystalline powder.

Compound 4:

NMR(CDCl₃) δ: 1.29(3H,t,J=7.0 Hz), 4.230(2H,q,J=7.0 Hz), 6.44(1H,bs),7.01(1H,bs), 7.59(2H,d,J=5.6 Hz), 8.60(2H,d,J=5.6 Hz) IR(Nujol) ν cm-1:1702, 1599

(3) To a solution of sodium metal (2.58 g, 0.11 moles) in 250 ml ofanhydrous ethanol was added a solution prepared by suspending 3 (15.58g, 51 mmol) in 70 ml of ethanol under ice cooling. The reaction washeated to reflux for 9 hours. The reaction solution was concentratedunder reduced pressure and the resulting residue was neutralized with 1Nhydrogen chloride. The precipitated insoluble materials were collectedby filtration, washed with water and then recrystallized fromisopropanol to obtain 3.0 g of 4 (yield 27.3%).

(1) To 4 (6.02 g, 27.8 mmol) were added 60 ml of ethanol and 60 ml of anaqueous solution containing NaOH (6.8 g, 0.17 moles), and the mixturewas heated to reflux for 1 hour. After ethanol in the reaction solutionwas distilled away under reduced pressure, the solution was neutralizedby adding acetic acid. The insoluble materials were collected byfiltration, washed with water and then dried to obtain 4.53 g of 5(yield 86.6%).

Compound 5: NMR(d₆-DMSO) δ: 6.42(1H,bs), 7.04(1H,bs), 7.56(2H,d,J=5.6Hz), 8.52(2H,d,J=5.6 Hz)

(2) To a solution of 5 (3.28 g, 17.4 mmol) in 30 ml of DMF was added H₂O(0.6 ml), and the mixture was heated to reflux overnight. The reactionwas concentrated under reduced pressure. The residue was recrystallizedfrom methanol to obtain 1.28 g of 6 (yield 51.0%).

Compound 6: NMR(d₆-DMSO) δ: 6.58(1H,bs), 6.86(1H,bs), 7.49(1H,bs),7.00(2H,d,J=6.2 Hz), 8.41(2H,d,J=6.2 Hz) IR(Nujol) ν cm-1:3144, 3104,3024, 1704, 1605

(1) To a solution of 7 (10.7 g, 0.1 moles) and 8 (22.4 g, 0.1 moles) in220 ml of THF were added H₂O (30 ml) and K₂CO₃ (16.6 g, 0.12 moles)successively, and the mixture was stirred at room temperature overnight.The reaction was concentrated under reduced pressure and the resultingresidue was dissolved in ethyl acetate. The solution was washed withwater, dried and then concentrated under reduced pressure. The residuewas recrystallized from ethyl ether-n-hexane to obtain 15.61 g of 9(yield 88.1%).

Compound 9: NMR(CDCl₃) δ : 1.35(3H,t,J=7 Hz), 4.28(2H,q,J=7 Hz),6.60(1H,d,J=16 Hz), 7.38(2H,d,J=6 Hz), 7.60(1H,d,J=16 Hz), 8.65(2H,d,J=6Hz) IR(CHCl₃) ν cm⁻¹1:1711, 1645, 1597, 1551

(2) To a suspension of t-BuOK (12.5 g, 0.106 moles) in 150 ml of THF wasadded dropwise a solution of 9 (15.61 g, 88 moles) and 10 (18.92 g, 97mmol) in THF (150 ml) over 40 minutes so as to keep the reactiontemperature below 30° C. The reaction solution was stirred at roomtemperature for 1 hour and neutralized with 10% HCl. The reactionsolution was distilled to remove THF and dissolved in ethyl acetate. Thesolution was washed with water, dried over anhydrous magnesium sulfateand concentrated under reduced pressure. The residue was purified bysilica gel column chromatography (ethyl acetate). The residue of theeluent was washed with ethyl acetate to obtain 13.9 g of 11 (yield73.0%). The corresponding carboxylic acid 11′ can be obtained bytreating the product according to the same manner as that described inPreparation 2.

To a suspension of LiAlH₄ (12.14 g, 0.32 moles) in 500 ml of THF wasadded slowly a solution of a compound 11 (34.55 g, 0.16 moles) in THF(500 ml), and the mixture was refluxed for 1 hour and 30 minutes andice-cooled. When a saturated Na₂SO₄ solution and 4N NaOH were added tothe mixture, oily insoluble materials were precipitated. The motherliquor was decanted and the insoluble materials were washed with THF.The mother liquors were combined and concentrated under reduced pressureand treated by silica gel column chromatography (methanol/ethylacetate=1/9). The eluent was concentrated under reduced pressure and theresidue was washed in turn with ethyl ether and hexane to obtain 21.01 gof 12 (yield 83%) as a pale yellow crystal. mp.152-155° C.

Compound 12: NMR(d₆-DMSO) δ: 2.22(3H,s), 6.66(1H,bs), 7.25(1H,bs),7.43(2H,d,J=6.0 Hz), 8.43(2H,d,J=6.0 Hz) IR(KBr) ν cm⁻¹: 3468, 1600

(1) To a solution of 7 (18.53 g, 173 mmol) in THF (250 ml) was addedPh₃P=COCH₃ (55 g, 173 mmol) at room temperature, and the mixture wasstirred for 1 hour and concentrated under reduced pressure. To theresidue were added toluene (50 ml) and hexane (20 ml). The precipitatedPh₃P═O was removed by filtration and the filtrate was concentrated toobtain 34.2 g of oily crude 13 (containing Ph₃P═O).

Compound 13: NMR(CDCl₃) δ: 2.44(3H,s), 6.86(1H,d,J=16.4 Hz),7.45(1H,d,J=16.4 Hz), 7.35(2H,dd,J=1.4,4 Hz), 8.69(2H,dd,J=1.4,4 Hz)

(2) To 90% t-BuOK (21.59 g, 173.17 mmol)/THF was added a mixture of thecrude 13 prepared in the above at 20° C. and tosylmethyl isocyanide(33.81 g, 173.17 mmol)/THF (300 ml) under ice cooling at 20 to 25° C.After stirring at 25° C. for 1 hour, acetic acid (0.5 ml) was added, andthen H₂O (300 ml) and ethyl acetate (700 ml) were added to separate theorganic layer. The organic layer was washed with water and concentratedunder reduced pressure. The residue was crystallized from toluene toobtain 21.38 g of a crude product 14. The yield from 7 was 66.4%. m.p.177-194° C.

Compound 14: NMR(d₆-DMSO) δ: 2.39(3H,s), 7.19(1H,bs),7.45(2H,dd,J=1.6,4.6 Hz), 7.80(1H,bs), 8.43(2H,dd,J=1.6,4.6 Hz)

(3) To an ethanol solution (200 ml) of 14 (20.9 g, 112.3 mmol) was addedNaBH₄ (4.25 g)/H₂O (15 ml), and the mixture was stirred at roomtemperature overnight. Excess NaBH₄ was decomposed with acetic acid. Thesolution was concentrated under reduced pressure. To the residue wasadded H₂O/ethyl acetate. The aqueous layer was made basic with K₂CO₃.The organic layer was separated, washed with a saturated saline solutionand concentrated. The residue was crystallized from toluene/ethylacetate (1/1) to obtain 15 (14.76 g). m.p. 153-156° C.

Compound 15: NMR(d₆-DMSO) δ: 1.38(3H,d,J=6.2 Hz), 4.78-4.95(1H,m),6.84(1H,bs), 7.20(1H,bs), 7.60(2H,dd,J=1.6,4.6 Hz), 8.43(2H,dd,J=1.6,4.6Hz) IR(Nujol) ν cm⁻¹: 3308, 1596, 1530, 1418, 1213, 1058, 979

(4) To 15 (9.03 g, 48.03 mmol) was added acetic anhydride (40 ml), andthe mixture was stirred at 90° C. for 1 hour. Excess acetic anhydridewas distilled away under reduced pressure. To the oily residue was addedH₂O (30 ml)/ethyl acetate (150 ml). K₂CO₃ was added until the solutionbecame basic. The organic layer was separated, washed with water andconcentrated under reduced pressure to obtain 15-1 (oily) (9.14 g).After dissolving 15-1 in THF (50 ml) and adding DBU (12 ml), thereaction was conducted at 70° C. for 8 hours. The reaction solution wasconcentrated under reduced pressure. The residue was dissolved in ethylacetate, washed with water and concentrated. To a solution of theresidue in methanol (30 ml) was added 2N NaOH (20 ml), and the mixturewas stirred at room temperature for 1 hour. After the completion of thereaction, the reaction solution was concentrated under reduced pressureuntil the volume becomes about 25 ml, and extracted with ethyl acetate.The residue was crystallized from toluene to obtain 16 (3.73 g, yield:45.7% from 15). m.p. 159-160° C.

Compound 16: NMR(d₆-DMSO) δ: 5.00(1H,dd,J=4.0,10.8 Hz),5.42(1H,dd,J=4,17.4 Hz), 6.70(1H,dd,J=10.8,17.4 Hz), 7.09(1H,bs),7.17(1H,bs), 7.36(1H,d,J=5.8 Hz), 8.47(1H,d,J=5.8) IR(Nujol)ν cm⁻¹:2716,1932, 1599, 1520, 1412, 1211, 1076, 986

(1) To a solution of 6 (1.44 g, 10 mmol) in 20 ml of DMF was added NaH(0.42 g, 10.5 mmol) under ice cooling, and the mixture was stirred atroom temperature for 30 minutes and cooled to −30° C. After addingchloromethyl benzyl ether (ClCH₂OCH₂Ph) (1.46 ml, 10.5 mmol) to thesolution, the mixture was stirred at the same temperature for 30minutes. The reaction solution was poured into ice-cold water, andextracted with ethyl acetate. The extract was washed with water, driedand concentrated under reduced pressure. The residue was purified bysilica gel chromatography (CH₂Cl₂/ethyl acetate=9/1-1/1) to obtain 2.31g of 18 (yield 87.4%).

Compound 18: NMR(CDCl₃) δ: 4.46(2H,s), 5.29(2H,s), 6.59(1H,bs),6.67(1H,bs), 7.26(1H,bs), 7.3˜7.4(5H,m), 7.41(2H,d,J=6 Hz),8.51(2H,d,J=6 Hz)

(2) To a solution of 18 (26.9 g, 0.1 moles) in 150 ml of DMF was addeddropwise phosphorous oxychloride (27.7 ml, 0.3 moles) while maintainingthe reaction temperature below 25° C., and the mixture was stirred withheating at 40 to 45° C. for 3 hours. The reaction solution was pouredinto 500 ml of water containing 127 g of K₂CO₃ under ice cooling. Thesolution was then extracted with ethyl acetate, washed with water, andconcentrated under reduced pressure. The resulting residue wascrystallized from ether to obtain 17.15 g of 19 (yield 60.0%).

Compound 19: NMR(CDCl₃) δ: 4.60(2H,s), 5.85(2H,s), 7.33(6H,m),41(2H,d,J=5.6 Hz), 7.54(1H,bs), 8.60(2H,d,J=5.6 Hz)

(3) To a solution of 19 (7.8 g, 27.7 mmol) in 150 ml of dichloromethanewas added 15 ml of anisole and aluminum chloride (11 g, 83.1 mmol) inseries at room temperature, and the mixture was stirred for 4 hours.After adding aluminum chloride (7.4 g, 55.4 mmol), the mixture wasstirred for 1 hour. The reaction solution was poured into ice-cold waterand diluted hydrochloric acid was added thereto to give a clearsolution. The aqueous solution was washed with ethyl ether, adjusted topH 8 with 4N NaOH and extracted with methyl ethyl ketone. The organiclayer was dried and concentrated under reduced pressure. The crystallineresidue was washed with ethyl ether-n-hexane to obtain 3.56 g of 20(yield 74.3%).

Compound 20: NMR(CD₃OD) δ: 7.51(1H,bs), 7.65(2H,d,J=6.0 Hz),7.79(1H,bs), 8.44(2H,d,J=6.0 Hz), 9.55(1H, bs) Compound 20: NMR(DMSO) δ:7.57(1H,bs), 7.64(2H,d,J=6.2 Hz), 7.97(1H,bs), 8.49(2H,d,J=6.2 Hz),9.55(1H, bs) IR(Nujol) ν cm⁻¹:1671, 1655, 1603

To a solution of 6 (5.77 g, 40 mmol) in 70 ml of DMF was added dropwisephosphorous oxychloride (14.5 ml, 0.116 moles) under ice cooling. Afterstirring at 130° C. for 5 hours, the solution was poured into ice-coldwater and neutralized with sodium hydrogencarbonate. To the solution wasadded ethyl acetate and the insoluble materials were removed byfiltration. The organic layer was washed with a saturated salinesolution, dried, and concentrated under reduced pressure. The residuewas recrystallized from methanol to obtain 0.97 g of 21 (yield 14.0%).

(1) To a solution of 12 (21.0 g, 0.13 moles) in 250 ml of DMF was added60% NaH (5.72 g, 0.143 moles) under ice cooling, and the mixture wasstirred at the same temperature for 20 minutes. To the reaction wasadded 100 ml of DMF, and the mixture was cooled to −45° C. After addingPOM-Cl (ClCH₂OCOBu^(t)) (20.6 ml, 0.143 moles) to the reaction solution,the mixture was stirred at the same temperature for 30 minutes, pouredinto H₂O (700 ml) and extracted twice with ethyl acetate. The ethylacetate layer was washed with water, a saturated saline solution, driedover MgSO₄, and distilled away under reduced pressure. The residue wastreated by silica gel chromatography (toluene/ethyl acetate=1/2) toobtain 33.73 g of a white crystal 22 (yield 95%). m.p. 42-44° C.

Compound 22: NMR(CDCl₃) δ: 1.19(9H,s), 2.24(3H,s), 5.75(2H,s),6.68(1H,d,J=2.4 Hz), 7.07(1H,d,J=2.4 Hz), 7.34(2H,d,J=6.2 Hz),8.53(2H,d,J=6.2 Hz) IR(CHCl₃) ν cm⁻¹:1732, 1602

(2) To 150 ml of DMF cooled at −20° C. was added POCl₃ (45.9 ml, 0.49moles). A solution of 22 (33.4 g, 0.12 moles) in DMF (40 ml) was thenadded dropwise. The solution was stirred at room temperature for 15minutes, and then at 60° C. for 80 minutes. The resulting reactionsolution was poured into 1000 ml of water, and K₂CO₃ (102 g, 0.74 moles)was added thereto. After adding 700 ml of ethyl acetate, the solutionwas neutralized with NaHCO₃. The ethyl acetate layer was washed withwater and a saturated saline solution, dried over MgSO₄, and distilledaway under reduced pressure. To the residue were added ethyl ether andhexane, and the precipitated crystals were filtered to obtain 27.6 g ofa pale yellow crystal 23 (yield 75%). m.p. 76-78° C.

Compound 23: NMR(CDCl₃:1.18(9H,s), 2.51(3H,s), 6.24(2H,s),7.29(2H,d,J=6.4 Hz), 7.33(1H,s), 8.62(2H,d,J=6.4 Hz), 9.90(1H,s)IR(CHCl₃) ν cm⁻¹:1731, 1657, 1603

To a solution of 23 (20.0 g, 66.6 mmol) in 500 ml of methanol was addeda solution of NaOH (13.3 g, 0.33 moles) in H₂O (200 ml), and the mixturewas stirred at room temperature for 1 hour. The reaction solution wasconcentrated under reduced pressure, and the precipitated insolublematerials were filtered to obtain 11.0 g of a white crystal 24 (yield89%). m.p. 214-216° C.

Compound 24: NMR(d₆-DMSO) δ: 2.50(3H,s), 7.50(2H,d,J=6.0 Hz),7.65(1H,bs), 8.53(2H,d,J=6.0 Hz), 9.73(1H,s), 12.24(H,bs) IR(KBr) νcm⁻¹: 1655, 1604

(1) To a solution of 20 (3.46 g, 20 mmol) in 20 ml of DMF was added(Boc)₂O (5.1 ml, 22 mmol), and the mixture was stirred at roomtemperature for 1 hour. The reaction solution was poured into ice-coldwater and extracted with ethyl acetate. The organic layer was washedwith water, dried and concentrated under reduced pressure. The resultingresidue was recrystallized from toluene to obtain 5.24 g of 25 (yield96.2%).

Compound 25: NMR(CDCl₃) δ: 1.68(9H,s), 7.43(2H,d,J=6.0 Hz),7.51(1H,d,J=2.0 Hz), 7.82(1H,d,J=2.0 Hz), 8.62(2H,d,J=6.0 Hz) IR(CHCl₃)ν cm⁻¹: 1755, 1666, 1604

(2) To a solution of 25 (2.72 g, 10 mmol) in 50 ml of methanol was addedO-methylhydroxylamine hydrochloride (0.92 g, 11 mmol), and the mixturewas stirred at room temperature for 1 hour. The reaction solution wasconcentrated under reduced pressure, and the resulting residue wasdissolved in water. The solution was neutralized with NaHCO₃, extractedwith ethyl acetate, washed with water, dried, and concentrated underreduced pressure to obtain 2.99 g of 26 (yield 92.2%) as a crystallineresidue.

Compound 26: NMR (CDCl₃ (mixture of syn/anti)) δ: 1.64, 1.65(9H,s+s),3.96/4.08=5/1(3H,s+s), 7.09/7.45=5/1(1H,bs+bs), 7.43(2H,d,J=6.2 Hz),7.68/7.72=1/5(1H,bs+bs), 8.58(2H,d,J=6.2 Hz), 8.28/8.64=1/5(1H,s)IR(CHCl₃) ν cm⁻¹: 11747, 1605

(3) To a suspension of 6.0 g of zinc powder in 20 ml of acetic acid wasadded dropwise a solution obtained by dissolving 26 (3.01 g, 10 mmol) in20 ml of acetic acid with vigorously stirring under ice cooling, whilemaintaining the reaction temperature below 30° C. After stirring for 30minutes, the zinc powder was removed by filtration and the mother liquorwas concentrated under reduced pressure. The residue was dissolved inwater and the solution was neutralized with NaHCO₃. To the solution wasadded aqueous ammonia, and the solution was extracted with ethylacetate. The organic layer was washed with water, dried, andconcentrated under reduced pressure to obtain 2.63 g of a crude product27. The crude product 27 was dissolved in 50 ml of THF, andcarbonyldiimidazole (2.43 g, 15 mmol) was added thereto. The mixture wasstirred at room temperature for 1 hour and concentrated under reducedpressure. The crystalline residue was washed with water and ethyl ether,and dried to obtain 3.02 g of 28 (yield 82.2%).

Compound 28: NMR(d₆-DMSO) δ: 1.58(9H,s), 4.66(2H,d,J=5 Hz), 6.85(1H,s),7.05(1H,s), 7.68(2H,d,J=6 Hz), 7.75(1H,s), 8.01(1H,s), 8.33(1H,s),8.50(2H,d,J=6 Hz), 9.93(1H,t,J=5 Hz) IR(Nujol) ν cm⁻¹:3183, 1746, 1704,1601

(4) To a solution of 28 (3.02 g, 8.22 mmol) and cyanamide (0.69 g, 16.4mmol) in 50 ml of DMF was added sodium hydride (0.33 g, 8.22 mmol), andthe reaction solution was stirred for 30 minutes with heating at 60° C.After adding 0.5 ml of acetic acid, the solution was concentrated underreduced pressure. To the residue were added ice-cold water and ethylether, and the mixture was adjusted to pH 6 by adding dilutedhydrochloric acid with stirring. The precipitated insoluble materialswere collected by filtration, washed with ethyl ether and water, anddried to obtain 1.37 g of 29 (yield 48.8%).

Compound 29: NMR(CDCl₃+CD₃OD) δ: 1.65(9H,s), 4.57(2H,s), 6.62(1H,d,J=2Hz), 7.43(2H,d,J=6 Hz), 7.60(1H,d,J=2 Hz), 8.50(2H,d,J=6 Hz) IR(CHCl₃) νcm⁻¹:2260, 2155, 1740, 1610 Elemental analysis for C₁₇H₁₉N₅O₃. 1.5H₂O:Calcd: C,55.43; H,6.02; N,19.01 Found: C,55.35; H,5.73; N,18.63

(1) To a solution of 20 (864 mg, 5 mmol) in 10 ml of formic acid wasadded hydroxylamine hydrochloride (417 mg, 6 mmol), and the mixture wasstirred at 110° C. for 6 hours. The reaction solution was concentratedunder reduced pressure. The residue was dissolved in water, and thesolution was neutralized with NaHCO₃. The precipitated crystals werecollected by filtration and recrystallized from methanol to obtain 394mg of 30 (yield 46.6%).

Compound 30: NMR(d₆-DMSO) δ: 7.56(1H,bs), 7.61(2H,d,J=6.2 Hz),7.91(1H,bs), 8.49(2H,d,J=6.2 Hz) IR(Nujol) ν cm⁻¹:3112, 2210, 1602, 1535

(2) To a solution of 30 (6.85 g, 40.5 mmol) in 500 ml of methanol wasintroduced hydrochloric acid gas under ice cooling until saturation isaccomplished. The solution was stirred at the same temperature for 1hour and then at room temperature for 1 hour. The reaction solution wasconcentrated under reduced pressure to obtain a crude product 31 as acrystalline residue. To the residue were added 100 ml of methanol and160 ml of a solution of ammonia (5.3 moles) in methanol, and the mixturewas stirred at room temperature for 24 hours. The reaction solution wasconcentrated under reduced pressure and the residue was dissolved in 150ml of water. The solution was neutralized with 2N NaOH. The precipitatedinsoluble crystals were collected by filtration, washed with water anddried to obtain 6.78 g of 32 (yield 89.9%).

Compound 32: NMR(d₆-DMSO) δ:7.34(1H,s), 7.34(2H,d,J=6 Hz), 7.49(1H,s),8.32(2H,d,J=6 Hz) IR(Nujol) ν cm⁻¹:1667, 1602, 1550

(3) To a suspension of 32 (931 mg, 5 mmol) in 10 ml of DMF was addedcarbonyldiimidazole (891 mg, 5.5 mmol) with stirring, and the mixturewas stirred at room temperature for 2 hours. Cyanamide monosodium saltwas prepared by dissolving cyanamide (0.42 g, 10 mmol) in 6 ml of DMF,adding NaH (0.4 g, 10 mmol) to the solution, and stirring the mixture atroom temperature for 30 minutes. The so obtained cyanamide monosodiumsalt was added to the above reaction mixture. The solution was stirredat room temperature for 3 hours, and at 50° C. for 1 hour. The reactionsolution was concentrated under reduced pressure. The resulting residuewas dissolved in water, neutralized by adding 10 ml of 1N hydrochloricacid, and the precipitated crystals were collected by filtration. Thecrystals were dissolved in 1N NaOH. A portion of insoluble materials wasremoved by filtration. The mother liquor was neutralized again withdiluted hydrochloric acid. The precipitated crystals were collected byfiltration, and dried to obtain 518 mg of 34 (yield 40.8%).

Compound 34: NMR(d₆-DMSO) δ: 7.58(2H,d,J=6 Hz), 8.06(1H,s), 8.11(1H,s),8.56(2H,bs) IR(Nujol) ν cm⁻¹:2180, 1626, 1597

(1) Compound 35 (14.32 g, 50 mmol) was dissolved in 240 ml of THF withheating. The solution was cooled to −70° C. and 100 ml of a solution ofMeMgBr (0.91 moles) in THF was added dropwise thereto at below −60° C.The solution was allowed to stand for 1 hour at the same temperature,and 20 ml of the same solution was added. After 1 hour, 100 ml of anaqueous solution containing 18 g of ammonium chloride was added to thereaction solution, followed by concentration under reduced pressure. Theresidue was extracted with ethyl acetate, washed with water andconcentrated under reduced pressure. The resulting residue wascrystallized from ethyl ether-n-hexane to obtain 14.0 g of 36 (yield86.5%).

Compound 36: NMR(CDCl₃) δ: 1.49(9H,s), 1.68(3H,d,J=7 Hz), 5.02(1H,q,J=7Hz), 5.97, 6.03(2H,ABq,J=8.0 Hz), 6.51(1H,d,J=2 Hz), 7.28(1H,d,J=2 Hz),7.35(2H,d,J=6 Hz), 8.48(2H,d,J=6 Hz

(2) To a solution of 36 (14.0 g, 46.3 mmol) in 250 ml of dichloromethanewas added 14 g of manganese dioxide, and the solution was heated toreflux with stirring for 1.5 hours. To the reaction was further added 7g of manganese dioxide five times every 1 hour. After adding 14 g ofmanganese dioxide, the solution was heated to reflux with stirringovernight. Further, 14 g of manganese dioxide was added and the solutionwas heated to reflux with stirring for 7 hours. Manganese dioxide wasremoved by filtration from the reaction solution. The solution waswashed with dichloromethane and methanol, and the mother liquor wasconcentrated under reduced pressure. The residue was recrystallized fromisopropanol to obtain 11.6 g of 37 (yield 83.4%).

Compound 37: NMR(CDCl₃) δ: 1.18(9H,s), 2.53(3H,s), 6.29(2H,s),7.30(1H,d,J=2 Hz), 7.40(2H,d,J=6.0 Hz), 7.51(1H,d,J=2 Hz),8.58(2H,d,J=6.0 Hz) IR(Nujol) ν cm⁻¹:1718, 1646, 1602

(3) To a solution of 37 (11.6 g, 39 mmol) in 120 ml of pyridine wasadded selenium dioxide (9.6 g, 86 mmol), and the solution was heated toreflux for 7 hours. The reaction solution was concentrated under reducedpressure and the residue was added to aqueous sodium hydrogencarbonate,and the mixture was stirred. The insoluble materials were removed byfiltration and treated with active carbon. The mother liquor wasadjusted to pH 5 with diluted hydrochloric acid. The precipitatedcrystals were collected by filtration, washed with water, and dried toobtain 8.05 g of 38 (yield 62.5%).

Compound 38: NMR(d₆-DMSO) δ: 1.11(9H,s), 6.21(2H,s), 7.71(2H,d,J=6.0Hz), 7.78(1H,d,J=1.6 Hz), 8.29(1H,d,J=1.6 Hz), 8.56(2H,d,J=6.0 Hz)

(4) To a solution of 38 (5.98 g, 18.1 mmol) in 100 ml of methanol wasadded 45 ml of 2N NaOH, and the mixture was stirred at room temperaturefor 2 hours. Methanol in the reaction solution was distilled away underreduced pressure, and the aqueous solution was neutralized by adding 18ml of 5N HCl. The precipitated crystals were collected by filtration anddried to obtain 4.8 g of 39.

Compound 2: NMR(d₆-DMSO) δ: 7.71(1H,bs), 7.79(2H,d,J=6.0 Hz),8.11(1H,bs), 8.55(2H,d,J=6.0 Hz)

(5) To a suspension of 39 (4.8 g, 22.2 mmol) in 150 ml of methanol and150 ml of dichloromethane was added diphenyldiazomethane (5.5 g, 28.3mmol), and the mixture was stirred at room temperature overnight. Thereaction solution was concentrated under reduced pressure, and theresulting residue was purified by silica gel column chromatography(dichloromethane/ethyl acetate=2/1) to obtain 3.92 g of 40 (yield56.6%). Compound 40: NMR(CDCl₃) δ: 7.12(1H,bs), 7.3˜7.5(12H,m),7.56(1H,bs), 7.63(1H,bs), 8.58(2H,d,J=6.0 Hz) IR(Nujol) ν cm⁻¹:3386,1725, 1645, 1603

(6) To a solution of 40 (2.72 g, 7.1 mmol) in 50 ml of dichloromethanewas added a solution of O-methylhydroxylamine hydrochloride (1.78 g,21.3 mmol) in methanol (15 ml), and the mixture was stirred at roomtemperature for 3 days. The reaction solution was concentrated underreduced pressure and the residue was neutralized with an aqueous sodiumhydrogencarbonate solution. Then, the insoluble crystal was collected byfiltration, washed with water and dried to obtain 2.93 g of 41.

Compound 41: NMR (d₆-DMSO(mixture of syn/anti) δ: 3.95/4.13:6.4/1(3H,2×s) 6.81(1H,bs), 7.12(1H,s), 7.3˜7.5(10H,m), 8.06(2H,d J=6 Hz),8.19(1H,bs), 8.71 (2H,d,J=6 Hz) IR(Nujol) ν cm⁻¹:3104, 2606, 1742, 1630,1600

(7) To a solution of 41 (0.82 g, 2 mmol) in 5 ml of dichloromethane wasadded 1 ml of anisole. After adding 6 ml of trifluoroacetic acid underice cooling, the mixture was stirred for 1 hour. The reaction solutionwas concentrated under reduced pressure and the residue was purified bycolumn chromatography (water-methanol) to obtain 0.65 g of 42.

Compound 42: NMR(d₆-DMSO) δ: 3.91(3H,s), 6.94(1H,bs), 7.42(1H,bs),7.9(3H,m), 8.55(2H,bs)

(8) To a solution of 42 (0.65 g, 2.65 mmol) in 10 ml of DMF was addedcarbonyldiimidazole (0.65 g, 4 mmol) under ice cooling, and the mixturewas stirred at room temperature for 2 hours. The reaction solution wasconcentrated under reduced pressure and the residue was dissolved inethyl acetate. The solution was washed with water, dried, andconcentrated under reduced pressure to obtain 586 mg of a crude product43 (yield 78.1%).

Compound 43: NMR (d₆-DMSO(mixture of syn/anti))δ:3.94/4.25=2.5/1(3H,s×2), IsomerA,3.94(3H,s), 6.65(1H,bs), 7.13(1H s),7.17(1H,bs), 7.35(2H,d,J=6 Mz), 7.53(1H,s), 8.02(1H,s), 8.52(2H d J=6Hz), IsomerB, 4.25(3H,s), 7.30(1H,bs), 7.41(1H,s), 7.42(2H,d,J=6 Hz),7.71(1H,bs), 7.73(1H s) 8.35(1H s) 8.57(2H d J=6 Hz)

(9) To a solution of 43 (586 mg, 2.1 mmol) in 4 ml of DMF was added asolution prepared by dissolving cyanamide (174 mg, 4.1 mmol) in DMF (2ml), adding sodium hydride (60%) (164 mg, 4.1 mmol) and stirring for 30minutes, and the mixture was stirred at room temperature for 1 hour. Tothe reaction solution was added 0.5 ml of acetic acid and the solutionwas concentrated under reduced pressure. The residue was eluted bycolumn chromatography (20% methanol-H₂O). The residue of the eluent waswashed with isopropanol to obtain 275 mg of a crystalline powder 44(yield 48.6%).

Compound 44: NMR(d₆-DMSO(single isomer)) δ: 3.82(3H,s), 6.88(1H,bs),7.98(1H,bs), 8.12(2H,d,J=6 Hz), 8.60(2H,d,J=6 Hz) IR(Nujol) ν cm⁻¹:3400,2180, 1675, 1640, 1620

(1) To a suspension of 5′ (3.07 g, 10 mmol) in 30 ml of DMF was added1-hydroxybenzotriazole (2.03 g, 15 mmol) and water-soluble carbodiimide(2.88 g, 15 mmol) in series, and the mixture was stirred at roomtemperature for 1 hour. To the reaction solution were added 45 (2.2 g,15 mmol) and a solution of diisopropylethylamine (2.6 ml, 15 mmol) inDMF (10 ml), and the mixture was stirred at room temperature for 1 hour.The reaction solution was poured into ice-cold water, and the solutionwas extracted with ethyl acetate, washed with water, dried and thenconcentrated under reduced pressure. The residue was crystallized fromethyl ether, and collected by filtration to obtain 3.23 g of 46 (yield81.9%).

Compound 46: NMR(CDCl₃) δ: 1.48(9H,s), 6.44(2H,s), 6.51(1H,bs),7.43(2H,d,J=6.2 Hz), 7.50(1H,d,J=2 Hz), 7.58(1H,d,J=2 Hz), 7.76 (1H,bs),8.56(2H,d,J=6.2H), 8.60(1H,bs) IR(Nujol) ν cm⁻¹:3574, 3398, 1735, 1631,1595

(2) To a solution of cyanamide (210 mg, 5 mmol) in 5 ml of DMF was addedsodium hydride (60%) (120 mg, 3 mmol), and the mixture was stirred atroom temperature for 30 minutes. After adding 46 (394 mg, 1 mmol), themixture was stirred at room temperature for 7 hours. To the reactionsolution was added 0.5 ml of acetic acid, and the solution wasconcentrated under reduced pressure. To the residue was added ice-coldwater. The insoluble materials were collected by filtration, washed withwater and ethanol, and dried to obtain 218 mg of 47 (yield 85.7%).

Compound 47: NMR(d₆-DMSO) δ: 7.55(2H,d,J=6 Hz), 7.81(1H,s), 7.93(1H,s),8.50(2H,d,J=6 Hz), 8.64(1H,bs), 9.12(1H,bs) IR(Nujol) ν cm⁻¹:3420, 3360,2240, 1690, 1650, 1610 Elemental analysis for C₁₂H₁₀N₆O . 0.4H₂O: Calcd:C,55.13; H,4.16; N,32.14 Found: C,55.22; H,4.24; N,31.82

(1) To a solution of 24 (5.61 g, 30.1 mmol) in 140 ml of THF were added(Boc)₂O (8.3 ml, 36.1 mmol) and 500 mg of DMA, and the mixture wasstirred at room temperature for 30 minutes. The reaction solution wasdistilled away under reduced pressure. To the residue were added ethylether and hexane, and the precipitated crystals were filtered to obtain8.07 g of a white crystal 48 (yield 95%). m.p. 105-107° C.

Compound 48: NMR(CDCl₃) δ: 1.66(9H,s), 2.48(3H,s), 7.30 (2H,d,J=6.2 Hz),7.52(1H,s), 8.65(2H,d,J=6.2 Hz), 10.47(1H,s) IR(CHCl₃) ν cm⁻¹:1745,1660, 1604

(2) To a solution of 48 (8.06 g, 28.2 mmol) in 120 ml of methanol wereadded pyridine (2.73 ml, 33.8 mmol) and MeONH₂.HCl (2.47 g, 29.6 mmol)in series, and the mixture was stirred at room temperature for 80minutes. The reaction solution was distilled away under reducedpressure, and 100 ml of ethyl acetate and 100 ml of H₂O were added toseparate the ethyl acetate layer. The ethyl acetate layer was washedwith water and a saturated saline solution, dried over MgSO₄, and thendistilled away under reduced pressure to obtain 8.08 g of a whitecrystal 4(yield 91%). m.p. 104-106° C.

Compound 49: NMR(CDCl₃) δ: 1.61(9H,s), 2.34(3H,s), 3.97(3H,s),7.32(2H,d,J=6.2 Hz), 7.44(1H,s), 8.61(1H,s), 8.61(2H,d,J=6.2 Hz)IR(CHCl₃) ν cm⁻¹:1740, 1604

(3) To a suspension of 6.0 g of zinc powder in 20 ml of acetic acid and10 ml of ethanol was added a solution of 49 (3.0 g, 9.51 mmol) in aceticacid (15 ml), and the mixture was stirred at room temperature for 30minutes, and at 35° C. for 30 minutes. The zinc powder was removed byfiltration and the mother liquor was concentrated under reducedpressure. Then, CHCl₃ and H₂O were poured into the residue. The pH ofthe solution was adjusted to 9 with aqueous ammonia and sodiumhydrogencarbonate. The CHCl₃ layer was taken, washed with a saturatedsaline solution, dried over MgSO₄ and then distilled away under reducedpressure to obtain a crude product 50. The crude product 50 wasdissolved in 90 ml of THF. To the solution was added carbonyldiimidazole(1.54 g, 9.50 mmol), and the mixture was stirred at room temperature for30 minutes. The resulting reaction solution was poured into water, andextracted twice with ethyl acetate. The ethyl acetate layer was washedwith water and a saturated saline solution, dried over MgSO₄, anddistilled away under reduced pressure. The residue was washed with ethylether to obtain 2.65 g of a white powder 51 (yield 73%).

Compound 51: NMR(d₆-DMSO) δ: 1.53(9H,s), 2.20(3H,s), 4.65(2H,bs),7.01(1H,s), 7.49(2H,d,J=6.4 Hz), 7.70(1H,s), 7.73(1H,s), 8.27(1H,s),8.57(2H,d,J=6.4 Hz)

(4) Compound 51 (2.65 g, 6.95 mmol) was dissolved in 30 ml of DMF.Separately, cyanamide (H₂NCN) (321 mg, 7.63 mmol) was dissolved in 20 mlof DMF and NaH (306 g, 7.63 mmol) was added thereto, and the mixture wasstirred at room temperature for 10 minutes. The resulting solution wasadded to the solution of 51 under ice cooling. After stirring at roomtemperature for 30 minutes, acetic acid (0.88 ml, 15.4 mmol) was added.The solution was distilled away under reduced pressure. To the residuewas added H₂O. After adding 13.48 ml of 2N HCl, the mixture was stirredunder ice cooling. The precipitated insoluble materials were collectedby filtration, washed with isopropanol (x2) and ethyl ether (x2) toobtain 965 mg of a yellow powder 52 (yield 54%).

Compound 52: NMR(d₆-DMSO) δ: 2.17(3H,s), 4.20(2H,bs), 7.20(1H,bs),7.37(1H,bs), 7.44(2H,d,J=6.4 Hz), 8.45(2H,d,J=6.4 Hz), 10.95(1H,bs)

(1) To a solution of 35 (4.30 g, 15.0 mg) in 120 ml of THF was added 35′(6.02 g, 180.0 mg), and then the mixture was refluxed for 2 hours and 20minutes. The reaction solution was distilled away under reducedpressure, and the residue was dissolved in 80 ml of ethanol. Afteradding 37.5 ml of 2N NaOH, the mixture was stirred at 50° C. for 1 hour.The resulting solution was adjusted to about pH3 by adding 37.5 ml of 2NHCl, and the solution was concentrated under reduced pressure untilmethanol was removed. To the concentrated suspension were added 200 mlof ethyl acetate and H₂O (50 ml), and the insoluble materials werefiltered off to obtain 2.82 g of a pale yellow crystal 53 (yield 88%).m.p. 272-274° C.

Compound 53: NMR(d₆-DMSO) δ: 6.22(1H,d,J=16.0 Hz), 7.11(1H,bs),7.41(1H,d,J=16.0 Hz), 7.58(2H,d,J=5.6 Hz), 7.79(1H,bs), 8.47(2H,bs),11.91(1H,bs), 12.11(1H,bs) IR(KBr) ν cm⁻¹:3431, 1670, 1609

(2) To a suspension of 53 (2.82 g, 13.2 mmol) in 50 ml of DMF was addedcarbonyldiimidazole (3.03 g, 15.8 mmol), and the mixture was stirred atroom temperature for 45 minutes.

The reaction solution was distilled away under reduced pressure and theresidue was washed with H₂O to obtain 3.19 g of a yellow crystal 54(yield 92%). m.p. 213-215° C.

Compound 54: NMR(d₆-DMSO) δ: 7.17(1H,bs), 7.33(1H,d,J=15.6 Hz),7.43(1H,bs), 7.59(2H,d,J=4.8 Hz), 7.81(1H,bs), 7.84(1H,d,J=15.6 Hz),8.00(1H,bs), 8.51(1H,bs), 8.52(2H,d,J=4.8 Hz), 12.13(1H,bs) IR(KBr) νcm⁻¹:1708, 1692, 1618, 1602

(3) Compound 54 (1.90 g, 7.19 mmol) was dissolved in 50 ml of DMF.Separately, H₂NCN (332 mg, 7.90 mmol) was dissolved in 30 ml of DMF, andNaH (316 mg, 7.90 mmol) was added thereto, and the mixture was stirredat room temperature for 10 minutes. To the resulting solution was addedthe solution of 54 under ice cooling, and the mixture was stirred atroom temperature for 20 minutes. To the resulting reaction solution wasadded acetic acid (0.91 ml, 15.8 mmol), and the mixture was distilledaway under reduced pressure. To the residue was added H₂O and 3.60 ml of2N HCl in series. The precipitated insoluble materials were filtered offto obtain 1.74 g of a yellow crystal 55 (yield 100%). m.p. ≧300° C.

Compound 55: NMR(d₆-DMSO) δ: 6.30(1H,d,J=15.6 Hz), 7.18(1H,bs),7.46(1H,d,J=15.6 Hz), 7.69(2H,d,J=6.4 Hz), 7.89(1H,bs), 8.51(2H,d,J=6.4Hz) IR(Nujol) ν cm⁻¹:2164, 1626

(4) To a suspension of 54 (3.19 g, 12.1 mmol) in 50 ml of DMF were addedBoc₂O (3.33 ml, 14.5 mmol) and 4-dimethylaminopyridine (147 mg, 1.20mmol), and the mixture was stirred at room temperature for 30 minutes.The resulting solution was distilled away under reduced pressure and theresidue was washed with ethyl ether to obtain 3.98 g of a pale yellowcrystal 56 (yield 90%). m.p. 172-175° C.

Compound 56: NMR(d₆-DMSO) δ: 1.65(9H,s), 7.17(1H,bs), 7.54(1H,d,J=15.2Hz), 7.72(2H,d,J=6.2 Hz), 7.91(1H,bs), 7.97(1H,bs), 8.28(1H,bs),8.57(1H,d,J=15.2 Hz), 8.61(2H,d,J=6.2 Hz), 8.68(1H,bs) IR(KBr) νcm⁻¹:1748, 1696, 1602

(5) Compound 56 (3.98 g, 10.9 mmol) was dissolved in 80 ml of DMF andthe solution was cooled to −20° C.

Separately, H₂NCN (551 mg, 13.1 mmol) was dissolved in 30 ml of DMF, andNaH (439 mg, 10.9 mmol) was added thereto, and the mixture was stirredat room temperature for 10 minutes. The resultant solution wasice-cooled and added to the solution of 56, and the mixture was stirredat −20° C. for 1 hour and 30 minutes. To the resulting solution wasadded 10.9 ml of 2N HCl and the mixture was distilled away under reducedpressure. To the residue was added H₂O, and NaHCO₃ (917 mg, 10.9 mmol)was added to the mixture. The insoluble materials were collected byfiltration, and washed with ethyl ether to obtain 3.31 g of a yellowcrystal 57 (yield 93%). m.p. 230-235° C.

Compound 57: NMR(d₆-DMSO) δ: 1.62(9H,s), 6.46(1H,d,J=16.2 Hz),7.59(1H,bs), 7.85(2H,bs), 8.21(1H,d,J=16.2 Hz), 8.24(1H,bs), 8.60(2H,bs)IR(Nujol) ν cm⁻¹:2148, 1746, 1631

(1) To a solution of 25 (20.90 g, 76.8 mmol) in 360 ml of ethanol wasadded NaBH4 (2.91 g, 76.8 mmol) at −25° C. After stirring at the sametemperature for 1 hour, the pH was adjusted to about 7 by adding 1N HCl.The reaction solution was poured into 500 ml of H₂O and the solution wasextracted twice with ethyl acetate. Then, the ethyl acetate layer waswashed with water and a saturated saline solution, dried over MgSO₄, anddistilled away under reduced pressure. The resulting reside was washedwith ethyl ether to obtain 18.3 g of 58 as a white crystal (yield 87%).m.p. 125-128° C.

Compound 58: NMR(d₆-DMSO) δ: 1.59(9H,s), 4.65(2H,d,J=5.6 Hz),5.15(1H,t,J=5.6 Hz), 6.75(1H,d,J=2.0 Hz), 7.63(2H,d,J=6.0 Hz),7.91(1H,d,J=2.0 Hz), 8.50(2H,d,J=6.0 Hz) IR(CHCl₃) ν cm⁻¹:3532, 1731,1605

(2) To a suspension of 58 (18.30 g, 66.7 mmol) in 90 ml of DMF was addedSOCl₂ (7.26 ml, 100 mmol) under ice cooling, and the mixture was stirredat room temperature for 1 hour and 30 minutes. The resulting suspensionwas diluted with ethyl ether. The insoluble materials were collected byfiltration and dissolved in 180 ml of DMF. To the solution were addedK₂CO₃ (36.9 g, 267 mmol), H₂O (30 ml), a solution of KCN (8.7 g, 133moles) in H₂O (15 ml) and tetrabutylammonium bromide (2.15 g, 6.7 mmol)under ice cooling, and the mixture was stirred at the same temperaturefor 24 hours. The reaction solution was poured into H₂O (600 ml) andextracted twice with ethyl acetate. The ethyl acetate layer was washedwith water and a saturated saline solution, dried over MgSO₄ anddistilled away under reduced pressure. The resulting residue was treatedby silica gel chromatography (toluene/ethyl acetate (2/1 to 1/3)) toobtain 6.83 g of a white crystal 59 (yield 36%) and 3.36 g of a whitecrystal 60 (yield 27%). The above product 59 (6.82 g, 24.07 mmol) and 60(2.59 g, 14.13 mmol) were combined, and dissolved in 170 ml of 2.96NHCl/methanol. The solution was refluxed for 24 hours. The resultingsolution was distilled away under reduced pressure and the pH wasadjusted to about 8 by adding H₂O (200 ml) and NaHCO₃. The solution wasextracted twice with ethyl acetate. The EtOAc layer was washed withwater and a saturated saline solution, dried over MgSO₄ and distilledaway under reduced pressure. The resulting residue was washed with ethylether to obtain 6.33 g of a white powder 61 (yield 77%).

Compound 59 m.p.126-129° C. NMR(d₆-DMSO) δ: 1.62(9H,s), 4.21 (2H,s),6.87(1H,d,J=2.0 Hz), 7.65(2H,d,J=6.2 Hz), 8.03(1H,d,J=2.0 Hz),8.52(2H,d,J=6.2 Hz) IR(KBr) ν cm⁻¹:2255, 2203, 1739, 1603

Compound 60 m.p.168-172° C. NMR(d₆-DMSO) δ: 3.98(2H,s), 6.52(1H,bs),7.47(1H,s), 7.48(2H,d,J=6.2 Hz), 8.41(2H,d,J=6.2 Hz) IR(KBr) νcm⁻¹:2258, 1604

Compound 61 m.p.147-150° C. NMR(d₆-DMSO) δ: 3.64(3H,s), 3.66(2H,s), 6.41(1H,bs), 7.40(1H,bs), 7.46(2H,d,J=6.0 Hz), 8.39(2H,d,J=6.0 Hz),11.13(1H,bs) IR(KBr) ν cm⁻¹:3446, 1731, 1602

(3) To a suspension of 61 (6.33 g, 29.27 mmol) in 60 ml of methanol wasadded 17.6 ml of 2N NaOH. After stirring at room temperature for 30minutes, the precipitated insoluble materials were filtered off toobtain 5.59 g of a white powder 62 (yield 85%). m.p. ≧300° C.

Compound 62 NMR(D₂O) δ: 3.54 (2H,s), 6.46(1H,bs), 7.38(1H,bs),7.55(2H,d,J=4.6 Hz), 8.36(2H,d,J=4.6 Hz) IR(KBr) ν cm⁻¹:1638, 1609, 1571

(4) Compound 62 (1.0 g, 4.46 mmol) was suspended in H₂O (20 ml). Thesuspension was neutralized with 1N HCl (4.46 ml) and distilled awayunder reduced pressure. The resulting residue was suspended in 20 ml ofDMF, and carbonyldiimidazole (868 mg, 5.35 mmol) was added thereto underice cooling, and the mixture was stirred at room temperature for 1 hour.Separately, H₂NCN (20 6 mg, 4.91 mmol) was dissolved in 15 ml of DMF,and NaH (196 mg, 4.91 mmol) was added thereto, and the mixture wasstirred at room temperature for 10 minutes. The resulting solution wasadded to the above solution under ice cooling, and the mixture wasstirred at room temperature for 1 hour. To the resulting solution wasadded 4.91 ml of 1N HCl was added. Then, the solution was adjusted toabout pH 7 with acetic acid and distilled away under reduced pressure.To the residue was added H₂O, and the precipitated insoluble materialswere filtered to obtain 663 mg of 63 as a deep green crystal (yield66%). m.p. 228-230° C.

Compound 63: NMR(d₆-DMSO) δ: 3.49(2H,s), 7.46(1H,bs), 7.60(1H,bs),7.71(2H,d,J=6.0 Hz), 8.47(2H,d,J=6.0 Hz) IR(Nujol) ν cm⁻¹:2138, 1631

(1) To 16 (3.32 g, 19.5 mmol)/DMF (33 ml) was added 60% NaH (0.86 g, 1.1eq.), and the mixture was stirred at room temperature for 20 minutes.After cooling the mixture to −30° C., Cl—CH₂OCOBu^(t) (3.0 ml, 1.06 eq.)was added. The mixture was stirred at −10 to 0° C. for 1 hour, pouredinto ice-cold water, extracted with ethyl acetate, and washed with H₂Oand a saturated saline solution. The residue was purified by silica gelchromatography to obtain 17 (oily, 5.96 g, 21.9 mmol, yield: 112%).

Compound 17: NMR(CDCl₃) δ: 1.196(9H,s), 5.10(1H,dd,J=1.8,10.8 Hz),5.45(1H,dd,J=1.8,17.6 Hz), 5.78(2H,s), 6.64(1H,dd,J=10.8,17.6 Hz),5.78(2H,s), 6.64(1H,dd,J=10.8,17.6 Hz), 7.00(2H,bs),7.31(2H,dd,J=1.6,4.6 Hz), 8.55(2H,dd,J=1.6,4.6 Hz) IR(CHCl₃) νcm⁻¹:1733, 1602

(2) POCl₃ (7.5 ml) was added to DMF (20 ml) at −20° C., and the mixturewas stirred at 0° C. for 20 minutes. To the solution was added 17 (5.90g)/DMF (30 ml), and the mixture was heated at 55° C. for 1.5 hours. Themixture was poured into ice-cold water. The solution was neutralizedwith K₂CO₃ and extracted with ethyl acetate. The residue wascrystallized from toluene to obtain 4.57 g of 64 (yield 78% from 16).m.p. 148-152° C.

Compound 64: NMR(d₆-DMSO) δ: 1.15(9H,s), 5.94(2H,s),6.45(1H,dd,J=7.8,15.6 Hz), 7.40(2H,dd,J=1.6,4.6 Hz), 7.43(1H,d,J=2.2Hz), 7.65(1H,d,J=15.6), 7.81(1H,d,J=2.2 Hz), 8.58(2H,dd,J=1.6,4.6 Hz),9.54(1H,d,J=7.8 Hz) IR(Nujol) ν cm⁻¹; 3020, 1717, 1649, 1598, 1519,1413, 1283, 1208, 1131, 960

(3) To a solution of NaClO₂ (1.49 g)/NH₂SO₃H (1.60 g)/H₂O (35 ml) wasadded 64 (2.24 g)/methanol (22 ml) at 5° C. The mixture was stirred at 5to 10° C. for 40 minutes. After adding Na₂SO₄ (4.14 g)/H₂O (25 ml), themixture was stirred at 10° C. for 20 minutes, and distilled away underreduced pressure to remove methanol. The solution was extracted withethyl acetate, washed with water and concentrated. The residue wascrystallized from CH₂Cl₂/toluene to obtain 65 (1.17 g, yield: 49.6%).m.p. 205-207° C.

Compound 65: NMR(d₆-DMSO) δ: 1.15(9H,s), 5.91(2H,s), 6.14(1H,d,J=15.6Hz), 7.27-7.40(3H,m), 7.51(1H,d,J=15.6 Hz), 7.67(1H,d,J=2 Hz),8.57(2H,s,J=5.8 Hz) IR(Nujol) ν cm⁻¹;3100, 2446, 1746, 1686, 1604, 1399,1279, 1267, 1109, 972

(4) Carbonyldiimidazole (0.67 g) was added to 65 (1.00 g)/DMF at roomtemperature, and the mixture was stirred at room temperature for 1 hour.After adding a solution of H₂NCN (212 mg)/60% NaH 8177 mg)/DMF (15 ml),the mixture was stirred at room temperature for 2 hours, and at 40° C.for 1 hour. After adding acetic acid (0.29 ml), the mixture wasconcentrated under reduced pressure until the volume becomes about 5 mland dissolved in H₂O (50 ml). To the mixture were added ethyl acetate(10 ml) and hexane (10 ml). When acetic acid was added to neutralize theaqueous layer, crystals were precipitated. The crystals were collectedby filtration, washed with H₂O, and ethyl acetate to obtain 0.91 g of 66as a crude crystal (yield 84.7%). m.p. 192-193° C.

Compound 66: NMR(d₆-DMSO) δ: 1.14(9H,s), 5.94(2H,s), 6.23(1H,d,J=15.6Hz), 7.32-7.45(3H,m), 7.61(1H,d,J=15.6 Hz), 7.68(1H,d,J=1.8 Hz),8.61(1H,bs) IR(Nujol) ν cm⁻¹;3108, 2224, 1742, 1687, 1373, 1174, 1120,978

(5) To a suspension of 66 (830 mg, 2.35 mmol) in 20 ml of methanol wasadded aqueous 2N NaOH (6 ml, 12 mmol) at room temperature, and themixture was stirred at room temperature for 3 hour. After adding 16 mlof 2N HCl under ice cooling, methanol was distilled away under reducedpressure. The precipitated solids were then collected by filtration andwashed in turn with H₂O and ethyl ether. The solid materials werefiltered, and dried to obtain 530 mg of a yellow powder 67 (yield 95%).

Compound 67: NMR(d₆-DMSO) δ: 11.79(1H, m), 8.58(2H, m), 7.69(1H,d,J=15.6Hz), 7.54(1H,s), 7.40(2H,d,J=5.7 Hz), 7.27(1H,s), 6.21(2H,d,J=15.6 Hz)IR(Nujol) ν cm⁻¹:3350, 3192, 2170, 1626, 1528, 1349, 1066

(1) To a suspension of 6 (50 g, 346.8 mmol) in 500 ml of anhydrous DMFwas added 60% NaH (15.2 g, 381.5 mmol) under ice cooling and under N₂gas flow, and the mixture was stirred at room temperature for 20minutes. After cooling to −30° C., Bu^(t)CO₂CH₂Cl (53 ml, 367.6 mmol)was slowly added dropwise over 35 minutes, and the mixture was stirredat −10° C. for 2 hours. The reaction solution was poured into 2.4 L ofice-cold water, and extracted with 2L of ethyl acetate. The organiclayer was washed in turn with water and a saturated saline solution,dried over MgSO₄ and concentrated under reduced pressure. Then, theprecipitated crystals were collected by filtration, washed withcyclohexane, and dried to obtain 83.24 g of a white crystal 6′ (yield93%). m.p. 96-99° C.

Compound 6′: NMR(CDCl₃) δ: 8.51(2H,dd,J=4.6,1.6 Hz),7.37(2H,dd,J=4.6,1.6 Hz), 7.28(1H,m), 6.89(1H,dd,J=2.8,2.2 Hz),6.55(1H,dd,J=2.8,1.8 Hz), 5.82(2H,s), 1.18(9H,s) IR(Nujol) ν cm⁻¹:1723,1597, 1132

(2) Anhydrous DMF (372 ml, 4.8 moles) was cooled to −20° C. under N₂ gasflow, and POCl₃ (120 ml, 1.282 moles) was added dropwise over 40minutes. To the mixture was added a raw material 6′ (82.8 g, 320.5moles), and the mixture was heated to 60° C., and stirred for 3 hours.After cooling to room temperature, the reaction solution was poured intoice-cold water (2L). The solution was adjusted to pH 8 by adding K₂CO₃(266 g, 1.923 moles) with stirring. The precipitated solid materialswere collected by filtration and dissolved in 2.2 L of methyl ethylketone. The solution was dried over MgSO₄ and concentrated under reducedpressure. The residue was recrystallized from toluene to obtain 65.69 gof a flesh crystal 75 (yield 72%). m.p. 183-186° C.

Compound 75: NMR(CDCl₃) δ: 9.69(1H,d,J=1.0 Hz), 8.59(2H,dd,J=4.5,1.7Hz), 7.60(1H,m), 7.40(2H,dd,J=4.6 Hz,1.6 Hz), 7.32(1H,d,J=2.0 Hz),6.28(2H,s), 1.17(9H,s) IR(Nujol) ν cm⁻¹:1718, 1655, 1600, 1426, 1141

(3) To a solution of NaClO₂ (18.16 g, 200.8 mmol) and H₂NSO₃H (19.50 g,200.8 mmol) in 1636 ml of H₂O was added dropwise a solution of 75 (23 g,80.334 moles) in methanol (400 ml), and the mixture was stirred underice cooling for 3 hours. After adding dropwise 250 ml of an aqueousNa₂SO₃ (50.62 g, 401.6 mmol) under ice cooling, the mixture was stirredfor additional 30 minutes and distilled under reduced pressure to removemethanol. The precipitated solids were collected by filtration and driedto obtain 21.14 g of a green powder 76′ (yield 21.14%). m.p. 241.5-243°C.

Compound 76′: NMR(d₆-DMSO) δ: 12.79(1H,bs), 8.50(2H,dd, J=4.6,1.5 Hz),7.99(1H,d,J=2 Hz), 7.61(2H,dd,J=4.6,1.6 Hz), 7.47(1H,d,J=2.0 Hz),6.21(2H,s), 1.11(9H,s) IR(Nujol) ν cm⁻¹:1734, 1686, 1610, 1195, 1129

(4) To a solution of NH₂CN (12.965 g, 308 mmol) in 130 ml of DMF wasadded NaH (11.719 g, 292.98 mmol) under ice cooling and under nitrogengas flow. After the completion of the addition, the reaction solutionwas cooled to room temperature and stirred until the evolution of ahydrogen gas is nearly terminated.

To a solution of 76′ (46.62 g, 154.2 mmol) in 470 ml of DMF was addedcarbonyldiimidazole (32.51 g, 200 mmol) at room temperature, and themixture was stirred for 1 hour. After ice cooling, the previouslyprepared Na⁺[NH-CN]⁻/DMF solution was added dropwise. The solution wascooled to room temperature and stirred for 1 hour. After adjusting thepH of the solution to about 7 by adding 5N HCl under ice cooling, DMFwas concentrated under reduced pressure. To the resultant residue wasadded 900 ml of ice-cold water. The precipitated solids were collectedby filtration, washed in turn with H₂O and ethyl ether, and dried toobtain 50 g of a yellowish green powder 78 (yield 99%). m.p. 253-256° C.

Compound 78: NMR(d₆-DMSO) δ: 8.64(2H,d,J=6.6 Hz), 8.09(1H,d,J=1.8 Hz),7.96(2H,d,J=6.6 Hz), 7.43(1H,d,J=1.8 Hz), 6.32(2H,s), 1.11(9H,s)IR(Nujol) ν cm⁻¹:2184, 2142, 1711, 1634, 1565, 1226, 1155

(5) To a mixture of 78 (50.2 g, 154 mmol) in 1 L of methanol was addeddropwise 2N NaOH (385 ml, 770 ml) at room temperature. After stirring atroom temperature for 1 hour, the solution was adjusted to pH 7 by adding385 ml of 2N HCl under ice cooling, and methanol was distilled awayunder reduced pressure. The precipitated solids were collected byfiltration, washed in turn with H₂O, isopropanol and ethyl ether, anddried to obtain 32 g of a white powder 71 (yield 100%). m.p. 268-270° C.(decomposition).

Compound 71: NMR(d₆-DMSO) δ: 12.2(1H,m), 8.58(2H,d,J=6.2 Hz),7.96(2H,d,J=6.8 Hz), 7.88(1H,m), 7.29(1H,s) IR(Nujol) ν cm⁻¹: 3320,3186, 3066, 2624, 2144, 1633, 1570, 1527, 1407, 1336, 1215, 1200

(6) To a suspension of 71 (32 g, 150 mmol) in 60 ml of anhydrousmethanol was added dropwise a solution of 1.14M CH₃ONa/methanol (135 ml,150 mmol) under ice cooling and under a nitrogen gas flow, and themixture was stirred under ice cooling for 10 minutes. After adding 350ml of isopropanol, methanol was distilled away under reduced pressure.The precipitated solids were collected by filtration, washed in turnwith isopropanol and ethyl ether, and dried to obtain 35.6 g of a paleyellow powder 68 (yield 99%).

Compound 68: NMR(d₆-DMSO) δ: 11.44(1H,m), 8.39(2H,d,J=5.8 Hz),7.53(2H,d,J=6.0 Hz), 7.39(1H,m), 6.91(1H,m) IR(Nujol) ν cm⁻¹:3324, 2714,2154, 1609, 1576, 1507, 1219, 1146, 1131

(1) To a suspension of 76′ (5.5 g, 18.19 mmol) in 110 ml of methanol wasadded dropwise aqueous 2N NaOH (45 ml, 90 mmol) at room temperature, andthe mixture was stirred at room temperature for 90 minutes. After adding2N HCl (45 ml, 90 mmol) under ice cooling, methanol was distilled awayunder reduced pressure. The precipitated solids were collected byfiltration, washed in turn with H₂O and ethyl ether, and dried to obtain3.26 g of a pale green crystal 76 (yield 95%). m.p. ≧300° C.

Compound 76: NMR(d₆-DMSO) δ: 12.15(1H,bs), 8.45(2H,m), 7.72(1H,m),7.62(2H,d,J=5.70 Hz), 7.29(1H,m), IR(Nujol) ν cm⁻¹:1629, 1561, 1529,1210

(2) To a solution of 76 (3.25 g, 17.28 mmol) in 40 ml of anhydrous DMFwas added carbonyldiimidazole (5.60 g, 34.56 mmol) at room temperatureunder N₂ gas flow, and the mixture was stirred for 2.5 hours. DMF wasdistilled away under reduced pressure. To the residue was added 100 mlof ice-cold water to precipitate solid materials. The solids werecollected by filtration, washed in turn with H₂O, isopropanol and ethylether, and dried to obtain 3.7 g of a pale yellow powder 77 (yield 90%).m.p. ≧300° C.

Compound 77: NMR(d₆-DMSO) δ: 12.81 Hz(1H,bs), 8.49˜8.54(3H,m),8.08(1H,s), 7.89(1H,s), 7.75˜7.78(3H,m), 7.18(1H,s) IR(Nujol) νcm⁻¹:1662, 1599, 1219

(3) Compound 77 (3.65 g, 15.32 mmol) was added to 70 ml of anhydrousDMF. To the mixture was added dropwise (Boc)₂O (7 ml, 30.64 mmol) atroom temperature under N₂ gas flow. After adding a very small amount of4-dimethylaminopyridine, the mixture was stirred at room temperature for70 minutes. The reaction solution was concentrated under reducedpressure and 200 ml of ethyl ether was added to the resulting residue.The resultant precipitates were collected by filtration, washed withether and dried to obtain 4.48 g of a pale yellow powder 79 (yield 86%).m.p. ≧300° C.

Compound 79: NMR(d₆-DMSO) δ: 8.57(2H,d,J=5.8 Hz), 8.37(2H,m),7.78(3H,m), 7.67(1H,s), 7.19(1H,s), 1.43(9H,s) IR(Nujol) ν cm⁻¹:1749,1726, 1603, 1239

(4) NH₂CN (1.557 g, 37.04 mmol) was added to 30 ml of DMF, and 60% NaH(1.235 g, 30.87 mmol) was added thereto under ice cooling and undernitrogen gas flow. The mixture was stirred at room temperature for 15minutes.

Compound 79 (10.445 g, 30.87 mmol) was added to 140 ml of DMF and cooledto −45° C. To the solution was added dropwise the previously preparedNa⁺[NH-CN]⁻/DMF solution under N₂ gas flow over 15 minutes. Afterstirring at −30° C. for 90 minutes, 1N HCl (62 ml, 62 mmol) was addedunder ice cooling and the solution was concentrated under reducedpressure. To the residue was added 400 ml of ice-cold water, and the pHwas adjusted to about 7 by adding NaHCO₃ (2.59 g, 30.87 mmol). Theprecipitated solids were collected by filtration, washed in turn withH₂O, isopropanol and ethyl ether, and dried to obtain 8.97 g of a paleyellow powder 69 (yield 93%). m.p. 257-260° C. (decomposition).

Compound 69: NMR(d₆-DMSO) δ: 8.67(2H,d,J=6.0 Hz), 8.28(1H,d,J=1.6 Hz),8.02(2H,d,J=6.2 Hz), 7.33(1H,d,J=1.6 Hz), 1.56(9H,s) IR(Nujol) νcm⁻¹:2150, 1741, 1675, 1523

(1) To a solution of NaClO₂ (9.95 g, 110 mmol) and H₂NSO₃H (10.68 g, 110mmol) in 250 ml of H₂O was added dropwise a solution of 80 (15.0 g, 50moles) in methanol (150 ml). After stirring under ice cooling for 40minutes, a solution of Na₂SO₃ (27.7 g, 220 mmol) in H₂O (150 ml) wasadded dropwise, and the mixture was stirred for additional 20 minutesunder ice cooling. The precipitated solids were washed with water anddissolved in methanol (400 ml)/ethyl acetate (300 ml). To the solutionwas added 200 ml of toluene, and the solution was concentrated underreduced pressure. The precipitated solids were collected by filtrationto obtain 10.5 g of a white powder 81 (yield 66.8%). m.p. 211-214° C.

Compound 81: NMR(d₆-DMSO) δ:12.83(1H,bs), 8.56(2H,dd,J=4.5 Hz,1.5 Hz),7.62(1H,s), 7.42(2H,m), 6.18(2H,s), 2.42(3H,s), 1.11(9H,s) IR(Nujol) νcm⁻¹:3136, 2372, 1725, 1677, 1606, 1420, 1260, 1242, 1130, 1112, 1020,963

(2) To a solution of NH₂CN (723 mg, 17.2 mmol) in 40 ml of anhydrous DMFwas added 60% NaH (670 mg, 16.63 mmol), and the solution was stirred for1 hour at room temperature until evolution of hydrogen gas wasterminated.

To a solution of a raw material 81 (3.63 g, 11.47 mmol) in 50 ml ofanhydrous DMF was added carbonyldiimidazole (2.23 g, 13.76 mmol) underN₂ gas flow, and the mixture was stirred at room temperature for 1 hour.After ice cooling, the previously prepared Na⁺[NH-CN]⁻/DMF solution wasadded dropwise, and the mixture was stirred for 5 hours at roomtemperature. The pH was adjusted to about 7 by adding 2N HCl under icecooling, and DMF was distilled away under reduced pressure. To theresulting residue was added 300 ml of ice-cold water. The precipitatedsolids were collected by filtration, washed in turn with H₂O and ethylether and dried to obtain 3.9 g of a white powder 82 (yield 99%). m.p.209-211° C.

Compound 82: NMR(d₆-DMSO) δ: 8.69(2H,d,J=6.2 Hz), 7.88(2H,d,J=6.8 Hz),7.78(1H,s), 6.26(2H,s), 2.50(3H,s), 1.11 (9H,s)

IR (Nujol) ν cm⁻¹: 2712, 2144, 1710, 1579, 1526, 1459, 1336, 1281, 1255,1151.

(3) To a suspension of 82 (3.9 g, 11.46 mmol) in 240 ml of methanol wasadded dropwise aqueous 2N NaOH (30 ml, 15 mmol) at room temperature tocompletely dissolve the suspension. After stirring at room temperaturefor 1 hour, the pH of the solution was adjusted to about 7 by adding 130ml of 2N HCl under ice cooling. Methanol was distilled away underreduced pressure. The precipitated solids were collected by filtration,washed in turn with H₂O, isopropanol and ethyl ether, and dried toobtain 2.19 g of a yellowish white powder 72 (yield 84%). m.p. 226-230°C.

Compound 72: NMR (d₆-DMSO) δ: 11.76 (1H, m), 8.62 (2H, d, J=5.7 Hz),7.93 (2H, d, J=6.0 Hz), 7.60 (1H, d, J=2.4 Hz), 2.59 (3H, s).

IR (Nujol) ν cm⁻¹: 3162, 2606, 2144, 1631, 1558, 1515, 1454, 1333, 1213,1153.

(4) To a suspension of 72 (12.63 g, 55.8 mmol) in 335 ml of anhydrousmethanol was added dropwise a solution of 1M CH₃ONa/methanol (51.3 ml,53.0 mmol) under ice cooling and under nitrogen gas flow, and themixture was stirred at room temperature for 10 minutes. To the mixturewas added 335 ml of isopropanol, and then a mixed solution of 100 ml ofisopropanol and 100 ml of ethyl ether, and the mixture was stirred atroom temperature for a while. The precipitated solids were collected byfiltration, washed with ethyl ether, and dried to obtain 13.61 g of apale yellow powder 70 (yield 99%).

Compound 70: NMR (d₆-DMSO) δ: 11.08 (1H, bs), 8.44 (2H, dd, J=4.5 Hz,1.5 Hz), 7.39 (2H, dd, J=4.35 Hz, 1.65 Hz), 7.03 (1H, d, J=3.3 Hz), 2.48(3H, s).

IR (KBr) ν cm⁻¹: 3434, 3375, 2153, 1606, 1563, 1473, 1426, 1413, 1339,1229, 1145, 835, 811.

Compound 72 (8.0 g, 35.4 mmol) was added to 190 ml of anhydrous DMF, and(Boc)₂O (34.8 g, 159.3 mmol) was added dropwise thereto at roomtemperature. After addition of 4-dimethylaminopyridine (0.86 g, 7.08mmol), the mixture was stirred at room temperature for 45 minutes. Thereaction solution was poured into a mixed solution of 400 ml of ethylacetate and 100 ml of H₂O with stirring under ice cooling. The aqueouslayer was taken and concentrated under reduced pressure. The resultingresidue was neutralized with diluted hydrochloric acid. The precipitatedinsoluble materials were filtered off to obtain 4.08 g of 83 (yield35%).

Compound 83: NMR (d₆-DMSO) δ: 8.65 (2H, m), 7.86 (1H, s), 7.70 (2H, d,J=5.2 Hz), 2.23 (3H, s), 1.54 (9H, s). IR (KBr) ν cm⁻¹: 3104, 2174,1754, 1639.

(1) To a solution of 75 (2.86 g, 10 mmol) in 50 ml of anhydrous toluenewas added 84 (4.97 g, 13 mmol), and the mixture was heated to reflux inan oil bath at 120° C. for 3.5 hours. Upon cooling to room temperature,crystals were precipitated. The crystals were collected by filtrationand washed with hexane to obtain 3.9 g of 85.

To a suspension of 85 in 80 ml of methanol was added 2N NaOH (24 ml, 48mmol) at room temperature, and the mixture was stirred for 90 minutes.Methanol was distilled away under reduced pressure. The residue waswashed twice with 100 ml of toluene. The aqueous layer was taken andadjusted to pH 7 by adding 148 ml of 1N HCl. The precipitated solidswere collected by filtration, washed in turn with H₂O and ethyl ether,and dried to obtain 1.20 g of a yellow powder 86 (yield 50%). m.p. ≧300°C.

Compound 86: NMR (d₆-DMSO) δ: 11.93 (1H, bs), 8.48 (2H, d, J=5.4 Hz),7.88 (1H, m), 7.85 (1H, s), 7.63 (2H, d, J=5.8 Hz), 7.53 (1H, s).

IR (Nujol) ν cm⁻¹: 2714, 1619, 1530, 1374, 1203, 927.

(2) To a solution of NH₂CN (302 mg, 7.17 mmol) in 6 ml of DMF was added60% NaH (268 mg, 6.69 mmol) under ice cooling and under nitrogen gasflow, and the mixture was stirred at room temperature for 2.5 hours toobtain a solution of Na⁺[NH—CN]⁻/DMF. To a suspension of 86 (1.19 g,4.78 mmol) in 15 ml of anhydrous DMF was added carbonyldiimidazole (1.0g, 6.21 mmol) at room temperature, and the mixture was stirred for 3hours. To the reaction solution was added dropwise the previouslyprepared Na⁺[NH—CN]⁻/DMF solution under ice cooling, and the mixture wasstirred at room temperature for 90 minutes. The reaction solution wasadjusted to about pH 7 by adding acetic acid under ice cooling andconcentrated under reduced pressure. To the residue was added 200 ml ofice-cold water. The precipitated solids were collected by filtration,washed in turn with H₂O, isopropanol and ethyl ether, and dried toobtain 928 mg of a yellowish green powder 73 (yield 73%). m.p. ≧300° C.

Compound 73: NMR (d₆-DMSO) δ: 12.10 (1H, bs), 8.64 (2H, d, J=6.4 Hz),8.11 (2H, d, J=7 Hz), 8.09 (1H, s), 7.75 (1H, s), 7.52 (1H, s).

IR (Nujol) ν cm⁻¹: 2718, 2152, 1627, 1572, 1526, 1374, 1206.

(1) To a suspension of 75 (2.86 g, 10 mmol) in a mixed solution of 120ml of toluene and 30 ml of THF was added 25 mmol of 87 at roomtemperature under nitrogen gas flow, and the mixture was heated toreflux at 100° C. for 8 hours. After cooling to room temperature, thereaction solution was concentrated under reduced pressure to obtain 3.7g of 88. To a mixture of 88 and 80 ml of methanol was added 2N NaOH (25ml, 50 mmol) at room temperature, and the mixture was stirred for 3hours at room temperature. Methanol was distilled away under reducedpressure, and the residue was washed three times with 100 ml of toluene.The aqueous layer was neutralized and adjusted to pH 7 with 2N HCl. Theprecipitated solids were collected by filtration, washed in turn withH₂O and ethyl ether, and dried to obtain 11.8 g of a yellow powder 89(yield 52%). m.p. ≧300° C.

Compound 89: NMR (d₆-DMSO) δ: 8.68 (2H, d, J=6.4 Hz), 8.26 (2H, d, J=6.8Hz), 8.17 (1H, s), 7.51 (1H, s), 7.29 (1H, s), 2.13 (3H, s).

IR (Nujol) ν cm⁻¹: 1681, 1620, 1571, 1375, 1313, 1198, 1162, 1013.

(2) To a solution of NH₂CN (266 mg, 6.33 mmol) in 10 ml of anhydrous DMFwas added 60% NaH (236 mg, 5.91 mmol) under ice cooling, and the mixturewas stirred at room temperature for 50 minutes to prepare a solution ofNa⁺[NH—CN]⁻/DMF. Compound 89 (963 mg, 4.22 mmol) was added to 15 ml ofanhydrous DMF, and carbonyldiimidazole (889 mg, 5.49 mmol) was addedthereto under N₂ gas flow, and the mixture was stirred at roomtemperature for 1 hour. To the solution was added dropwise thepreviously prepared Na⁺[NH—CN]⁻/DMF solution under ice cooling, and themixture was stirred at room temperature for 2.5 hours. After addingacetic acid (0.72 ml, 12.66 mmol) under ice cooling, the solution wasconcentrated under reduced pressure. To the residue was added 200 ml ofice-cold water to precipitate. The precipitated solids were thencollected by filtration, washed in turn with H₂O, isopropanol and ethylether, and dried to obtain 1.05 g of a yellow powder 74 (yield 99%).

Compound 74: NMR (d₆-DMSO) δ: 12.44 (1H, bs), 8.73 (2H, d, J=6.2 Hz),8.29 (2H, d, J=6.8 Hz), 8.25 (1H, m), 7.40 (2H, s), 2.17 (3H, s).

IR (Nujol) ν cm⁻¹: 3272, 2718, 2240, 2148, 1670, 1596, 1523, 1374, 1323,1206, 1193, 1015.

(1) To a suspension of 91 (2.2 g, 7.32 mmol) in 40 ml of methanol wasadded aqueous 2N NaOH (18 ml, 36 mmol) at room temperature, and themixture was stirred for 1 hour. After adding dropwise 2N HCl (18 ml, 36mmol) under ice cooling, methanol was distilled away under reducedpressure. The residue was extracted three times with 50 ml of ethylacetate. Organic layers were combined and dried over MgSO₄. MgSO₄ wasremoved by filtration and the filtrate was concentrated under reducedpressure. The precipitated crystals were collected by filtration, washedwith ethyl ether, and dried to obtain 1.08 g of a white crystal 92(yield 79%). m.p. 179-181° C.

Compound 92: NMR (CDCl₃) δ: 9.60 (1H, bs), 8.58 (2H, d, J=5.4 Hz), 7.45(1H, dd, J=3.15, 1.65 Hz), 7.41 (2H, dd, J=6.3, 3.0 Hz), 7.23 (1H, dd,J=2.55, 1.65 Hz), 2.51 (3H, s).

IR (CHCl₃) ν cm⁻¹: 3438, 3002, 1650, 1602, 1391, 1313, 1273, 943.

(2) Compound 92 (1.08 g, 5.80 mmol) was suspended in 20 ml of anhydrousDMF under nitrogen gas flow. After adding dropwise (Boc)₂O (3.3 ml, 14.5mmol), 4-dimethylaminopyridine (71 mg, 0.58 mmol) was added to thesuspension, and the mixture was stirred at room temperature for 80minutes. The resulting solution was concentrated under reduced pressure.To the residue was added 60 ml of n-hexane, and the mixture was slowlystirred at room temperature. The precipitated crystals were thencollected by filtration, and dried to obtain 1.39 g of a whitish pinkcrystal 93 (yield 84%). m.p. 113-115° C.

Compound 93: NMR (CDCl₃) δ: 8.60 (2H, d, J=6.0 Hz), 7.71 (1H, d, J=1.8Hz), 7.39 (2H, dd, J=4.6, 1.6 Hz), 7.15 (1H, d, J=1.8 Hz), 2.52 (3H, s),1.61 (9H, s).

IR (CHCl₃) ν cm⁻¹: 2982, 1752, 1677, 1604, 1392, 1281, 1237, 1148.

(3) NaH (1.128 g, 28.20 mmol) was added to 30 ml of anhydrous THF. After(EtO)₂P(O)CH₂CO₂Et (5.73 ml, 28.92 mmol) was added dropwise at 25° C.,the mixture was stirred at room temperature for 20 minutes. Under icecooling, a solution of 93 (1.38 g, 4.82 mmol) in anhydrous THF (13 ml)was added dropwise over 10 minutes, and the mixture was stirred at roomtemperature for 100 minutes. Under ice cooling, 28 ml of 1N HCl wasadded and the solution was extracted three times with 50 ml of ethylacetate. The organic layers were combined, washed twice with a salinesolution, dried and concentrated. The resulting residue was purified bysilica gel column chromatography to obtain 433 mg of a white crystal 94(E-form) and 1.01 g of an orange oily substance 94 (Z-form), that is,total of 1.444 g (total yield: 84%).

E-form: m.p. 91-93° C.

NMR (CDCl₃) δ: 8.57 (2H, d, J=5.4 Hz), 7.68 (1H, dd, J=1.7, 1.1 Hz),7.38 (2H, dd, J=4.8 Hz, 1.2 Hz), 6.51 (1H, dd, J=1.9, 0.9 Hz), 5.99 (1H,s), 4.22 (2H, q, J=7.1 Hz), 2.42 (3H, s), 1.60 (9H, s), 1.32 (3H, t,J=7.2 Hz).

IR (CHCl₃) ν cm⁻¹: 2980, 1747, 1705, 1604, 1355, 1287, 1155, 1140.

Z-form:

NMR (CDCl₃) δ: 8.54 (2H, d, J=6.2 Hz), 7.71 (1H, dd, J=2.0, 0.8 Hz),7.38 (2H, dd, J=4.5, 1.7 Hz), 6.38 (1H, dd, J=1.9, 0.7 Hz), 5.98 (1H,dd, J=1.4, 0.8 Hz), 4.01 (2H, q, J=6.4 Hz), 2.21 (3H, s), 1.57 (9H, s),1.12 (3H, t, J=7 Hz).

IR (CHCl₃) ν cm⁻¹: 2976, 1743, 1710, 1604, 1370, 1277, 1220.

(4) To a solution of 94 (E) (334 mg, 0.94 mmol) in 7 ml of methanol wasadded dropwise aqueous 1N NaOH (4.7 ml, 4.7 mmol) at room temperature,and the mixture was stirred at 60° C. for 2 hours. Under ice cooling,14.7 ml of 1N HCl was added and methanol was distilled away underreduced pressure. The precipitated solids were collected by filtration,washed in turn with H₂O and ethyl ether, and dried to obtain 180 mg of apale yellow powder 95 (E) (yield 94%). m.p. 243-245° C.

Compound 95(E): NMR (d₆-DMSO+D₂O+DCl) δ: 12.35 (1H, bs), 8.66 (2H, d,J=6.9 Hz), 8.20 (2H, d, J=7.2 Hz), 8.12 (1H, d, J=0.9 Hz), 7.40 (1H, s),6.26 (1H, s), 2.48 (3H, s).

IR (Nujol) ν cm⁻¹: 3242, 1670, 1610, 1570, 1312, 1216, 1160, 1010, 799.

(5) To a solution of NH₂CN (249 mg, 5.92 mmol) in 8 ml of DMF was added60% NaH (304 mg, 7.6 mmol) under ice cooling, and the mixture wasstirred at room temperature for 40 minutes to prepare a solution ofNa⁺[NH—CN]⁻/DMF.

To a suspension of 95 (E) (169 mg, 0.74 mmol) in 7 ml of anhydrous DMFwas added carbonyldiimidazole (156 mg, 0.96 mmol) at room temperature,and the mixture was stirred at room temperature for 1 hour. To thesolution was added dropwise the previously prepared Na⁺[NH—CN]⁻/DMFsolution under ice cooling, and the mixture was stirred at roomtemperature for 6 hours. After adding dropwise acetic acid (0.97 ml,22.8 mmol) under ice cooling, the solution was concentrated underreduced pressure. To the residue was added 80 ml of ice-cold water, andthe mixture was stirred. The precipitated solids were collected byfiltration, washed in turn with H₂O, isopropanol and ethyl ether, anddried to obtain 128 mg of a yellow powder 96 (E) (yield 66%). m.p.217-220° C.

Compound 96(E): NMR (d₆-DMSO+DCl, D₂O) δ: 12.57 (1H, bs), 8.70 (2H, d,J=6.8 Hz), 8.23 (2H, d, J=6.8 Hz), 8.20 (1H, s), 7.49 (1H, d, J=0.4 Hz),6.28 (1H, s), 2.52 (3H, s).

IR (Nujol) ν cm⁻¹: 2142, 1629, 1537, 1300, 1255, 1202, 930.

(6) To a solution of 94 (Z) (517 mg, 1.45 mmol) in 11 ml of ethanol wasadded dropwise aqueous 1N NaOH (7.25 ml, 7.25 mmol) at room temperature,followed by stirring at 60° C. for 45 minutes. Under ice cooling, 17.25ml of 1N HCl was added, and ethanol was distilled away under reducedpressure. The precipitated solids were collected by filtration, washedin turn with H₂O and ethyl ether, and dried to obtain 250 mg of a paleyellow crystal 9 (Z) (yield 76%). m.p. 153-156° C.

Compound 95(Z): NMR (d₆-DMSO) δ: 13.23 (1H, s), 8.52 (2H, d, J=4.6 Hz),7.91 (1H, d, J=1.6 Hz), 7.73 (2H, d, J=5.8 Hz), 7.27 (1H, s), 5.69 (1H,d, J=0.8 Hz), 2.29 (3H, s).

IR (Nujol) ν cm⁻¹: 3368, 2156, 1630, 1558, 1507, 1208, 1154, 929.

(7) 60% NaH (76 mg, 1.887 mmol) was added to 2 ml of anhydrous DMF. Tothe mixture was added NH₂CN (86 mg, 2.04 mmol) under ice cooling andunder N₂ gas flow, and the mixture was stirred at room temperature for90 minutes to prepare a solution of Na⁺[NH—CN]⁻/DMF.

To a suspension of 95 (Z) (233 mg, 1.02 mmol) in 5 ml of anhydrous DMFwas added carbonyldiimidazole (255 mg, 1.632 mmol) at room temperature,and the mixture was stirred at room temperature for 105 minutes. To thesuspension was added dropwise the previously prepared Na⁺[NH—CN]⁻/DMFsolution under ice cooling, and the mixture was stirred at roomtemperature for 2 hours. After adding dropwise 1N HCl (13.6 ml, 3.6mmol) under ice cooling, DMF was distilled away under reduced pressure.To the residue was added 100 ml of ice-cold water. The precipitatedsolids were collected by filtration, washed in turn with H₂O,isopropanol and ethyl ether, and dried to obtain 174 mg of a brownpowder 96 (Z) (yield 68%). m.p. 219-221° C.

Compound 96(Z): NMR (d₆-DMSO) δ: 12, 29 (1H, bs), 8.58 (2H, d, J=6.2Hz), 7.99 (1H, s), 7.96 (2H, d, J=6.2 Hz), 7.19 (1H, s), 5.67 (1H, d,J=0.4 Hz), 2.21 (3H, s).

IR (Nujol) ν cm⁻¹: 2718, 2150, 1607, 1562, 1503, 1283, 1199, 1153.

(1) Compound 1 (23.72 ml, 214.4 mmol) was added to 215 ml of THF and themixture was cooled to −72° C. Under N₂ gas flow, (CO₂Et)₂ (29.12 ml,214.4 mmol) and a solution of LiN(TMS)₂/THF (1 mole, 214 ml) were addedin series, and the mixture was stirred while slowly elevating thetemperature up to room temperature. After cooling again to −65° C., 64ml of 5N HCl was added dropwise, and the mixture was stirred at roomtemperature for 30 minutes. The reaction solution was concentrated underreduced pressure. To the residue was added 128.7 ml of 1N NaOH. Theprecipitated crystals were collected by filtration, washed in turn withH₂O and ethyl ether, and dried to obtain 64 g of a yellow crystal 99.After cooling to −30° C., the crystals were dissolved in 126 ml ofconcentrated HCl and 51.5 ml of H₂O. To the solution was addedH₂NNH₂.H₂O (9.0 ml, 185 mmol), and the mixture was stirred at 85° C. for40 minutes. Then, 130 ml of 1N NaOH was added under ice cooling. Theprecipitated crystals were collected by filtration, washed in turn withH₂O, isopropanol and ethyl ether, and dried to obtain 27.9 g of a yellowpowder 100 (yield 70%). m.p. 215-217° C.

100: NMR (d₆-DMSO) δ: 14.4 (bs, 1H), 8.67 (2H, d, J=6 Hz), 7.95 (2H, d,J=6 Hz), 7.56 (1H, m), 4.35 (2H, q, J=7 Hz), 1.33 (3H, t, J=7.1 Hz).

IR (Nujol) ν cm⁻¹: 1726, 1609, 1572, 1248, 1204.

(2) To a suspension of 100 (2.17 g, 10 mmol) in 40 ml of ethanol wasadded dropwise 50 ml of 1N NaOH at room temperature, and the mixture wasstirred at 60° C. for 2 hours. Under ice cooling, 50 ml of 1N HCl wasadded. Methanol was distilled away under reduced pressure. Theprecipitated crystals were collected by filtration, washed in turn withH₂O, THF and ethyl ether, and dried to obtain 1.47 g of a white powder101 (yield 78%). m.p. ≧300° C.

Compound 101: NMR (d₆-DMSO+DCl) δ: 8.91 (2H, m), 8.50 (2H, d, J=5.8 Hz),7.80 (1H, s).

IR (Nujol) ν cm⁻¹: 3144, 2454, 2068, 1637, 1598, 1552, 1403, 1375, 1202,831, 807.

(3) NH₂CN (625 mg, 14.86 mmol) was added to 17 ml of anhydrous DMF. Tothe mixture was added 60% NaH (565 mg, 14.12 mmol) under ice cooling andunder N₂ gas flow, and the mixture was stirred at room temperature for 1hour to prepare a solution of Na⁺[NH—CN]⁻/DMF. To a suspension of 101(1.406 g, 7.432 mmol) in 50 ml of anhydrous DMF was addedcarbonyldiimidazole (2.049 g, 12.63 mmol) at room temperature under N₂gas flow, and the mixture was stirred for 2 hours. To the suspension wasadded dropwise the previously prepared Na⁺[NH—CN]⁻/DMF solution underice cooling, and the mixture was stirred at room temperature for 1 hour.After adding 120 ml of 1N HCl under ice cooling, DMF was distilled awayunder reduced pressure. To the residue was added 200 ml of ice-coldwater. The precipitated solids were collected by filtration, washed inturn with H₂O, isopropanol and ethyl ether, and dried to obtain 1.225 gof a white powder 102 (yield 77%).

m.p. ≧300° C.

Compound 102: NMR (d₆-DMSO, DCl) δ: 8.98 (2H, d, J=6 Hz), 8.52 (2H, d,J=6.2 Hz), 7.95 (1H, s).

IR (Nujol) ν cm⁻¹: 2170, 1635, 1573, 1541, 1345, 957.

(4) To a suspension of 102 (479 mg, 2 mmol) in 5 ml of anhydrousmethanol was added dropwise a solution of MeONa/methanol (1.1 M, 1.75ml) under ice cooling and under N₂ gas flow, and the mixture was stirredat 0° C. for 20 minutes. The reaction solution was poured into 50 ml ofisopropanol. The solution was concentrated under reduced pressure untilmethanol was removed. The solids were collected by filtration, washed inturn with isopropanol and ethyl ether, and dried to obtain 420 mg of awhite powder 103 (yield 89%).

Compound 103: NMR (D₂O) δ: 8.52 (2H, m), 7.67 (2H, m), 7.06 (1H, s).

IR (Nujol) ν cm⁻¹: 3090, 2150, 1611, 1581, 1562, 1413, 1375, 1340, 988.

(1) To a suspension of 104 (48 g, 276.5 mmol) in 500 ml of anhydrousethanol was added dropwise SOCl₂ (40.34 ml, 553 mmol) over 20 minutesunder ice cooling and under N₂ gas flow. After stirring at 50° C. for 70minutes, the solution was concentrated under reduced pressure. To theresidue were added 500 ml of ethyl acetate and 100 ml of H₂O, and the pHof the solution was adjusted to about 7 with Na₂CO₃. The ethyl acetatelayer was taken, concentrated under reduced pressure, and purified bydistillation under reduced pressure to obtain 44.96 g of 105 (yield98%).

Compound 105: NMR (CDCl₃) δ: 8.57 (2H, dd, J=6.0, 1.5 Hz), 7.25 (2H, d,J=6.0 Hz), 4.18 (2H, q, J=7.2 Hz), 3.62 (2H, s), 1.27 (3H, t, J=7.2 Hz).

(2) Compound 105 (44.83 g, 271.4 mmol) was added to 450 ml of anhydrousTHF, and HCO₂Et (65.77 ml, 814 mmol) was added to the mixture under N₂gas flow. After adding 60% NaOH (13.03 g, 325.68 mmol), the mixture wasstirred at room temperature for 3 hours. To the mixture was added 720 mlof ethyl ether. The resultant precipitates were collected by filtrationand dried to obtain 59.39 g of a powder 106.

Compound 106: NMR (D₂O) δ: 8.94 (1H, s), 8.36 (2H, d, J=6.2 Hz), 7.43(2H, d, J=6.2 Hz), 4.16 (2H, q, J=7.0 Hz), 1.25 (3H, t, J=7.3 Hz).

(3) Compound 106 (59.39 g, 276 mmol) was added to 480 ml of anhydrousTHF. To the mixture was added dropwise MsCl (21.36 ml, 276 mmol) underice cooling and under N₂ gas flow, and the mixture was stirred for 35minutes. To the mixture were added dropwise HSCH₂CO₂ET (30.26 ml, 276mmol) and Et₃N (42.32 ml, 303.6 mmol) in series, and the mixture wasstirred under ice cooling for 20 minutes. Then, 600 ml of ethyl acetateand 600 ml of H₂O were added. The ethyl acetate layer was taken, washedin turn with H₂O and a saturated saline solution, and dried over MgSO₄.MgSO₄ was removed by filtration, and the filtrate was concentrated underreduced pressure. The resulting residue was subjected to silica gelcolumn chromatography. The fraction eluted with toluene/ethyl acetate(2/1) was concentrated under reduced pressure to obtain 71.11 g of anoily substance 107 (yield 87%).

Compound 107: NMR (CDCl₃) δ: 8.64 (2H, dd, J=6.0, 1.6 Hz), 7.98 (1H, s),7.25 (2H, dd, J=6.0, 1.8 Hz), 4.24 (4H, q, J=7.0 Hz), 3.53 (2H, s), 1.31(3H, t, J=7.0 Hz), 1.29 (3H, t, J=7.1 Hz).

IR (CHCl₃) ν cm⁻¹: 2980, 1737, 1700, 1600, 1575, 1365, 1295, 1240, 1180.

(4) Compound 107 (71.11 g, 240.8 mmol) was added to 700 ml of anhydrousTHF, and the solution was cooled to −50° C. Under N₂ gas flow, asolution (265 ml, 265 mmol) of LiN(TMS)₂/THF (1 mole) was addeddropwise, and the mixture was stirred at room temperature for 1 hour.Acetic acid (30.33 ml, 529 mmol) was then added, and the solution wasconcentrated under reduced pressure. To the residue were added 1L ofethyl acetate and 500 ml of H₂O. The ethyl acetate layer was taken,washed in turn with H₂O and a saturated saline solution, and dried overMgSO₄. After removing MgSO₄ by filtration, the filtrate was concentratedunder reduced pressure. To the residue was added 300 ml of CH₂Cl₂ andthe insoluble materials were removed by filtration. The filtrate wasconcentrated under reduced pressure to obtain 56.82 g of 108 (yield95%). m.p. 123-125° C.

Compound 108: NMR (CDCl₃) δ: 10.28 (1H, bs), 8.65 (2H, dd, J=6.2, 1.8Hz), 7.68 (1H, s), 7.67 (2H, dd, J=6.4, 1.6 Hz), 4.42 (2H, q, J=7.0 Hz),1.41 (3H, t, J=7.2 Hz).

IR (CHCl₃) ν cm⁻¹: 2980, 1658, 1605, 1570, 1440, 1377, 1328, 1190.

(5) Compound 108 (56.82 g, 228 moles) was dissolved in 1000 ml ofanhydrous CH₂Cl₂ and the solution was cooled to −30° C. Under N₂ gasflow, (2,6-di-tert-butyl-4-methylpyridine) (51.50 g, 251 mmol) and(CF₃SO₂)₂O (47.95 ml, 285 mmol) were added dropwise in series, and themixture was stirred under ice cooling for 50 minutes. The reactionsolution was concentrated under reduced pressure. To the residue wereadded 1000 ml of ethyl acetate and 550 ml of saturated NaHCO₃. The ethylacetate layer was taken, washed in turn with H₂O and a saturated salinesolution, and dried over MgSO₄. MgSO₄ was removed by filtration and thefiltrate was concentrated under reduced pressure, and subjected tosilica gel column chromatography. The fraction eluted with toluene/ethylacetate (4/1) was concentrated under reduced pressure to obtain 67.32 gof 109 (yield 77%). m.p. 100-101° C.

Compound 109: NMR (CDCl₃) δ:8.72 (2H, dd, J=6.0, 1.4 Hz), 7.64 (1H, s),7.39 (2H, dd, J=6.2, 1.6 Hz), 4.46 (2H, q, J=6.8 Hz), 1.43 (3H, t, J=7.0Hz).

IR (CHCl₃) ν cm⁻¹: 2970, 1715, 1600, 1430, 1375, 1290, 1265, 1240, 1220.

(6) Pd(PPh₃)₄ (4.09 g, 3.54 mmol) and LiCl (22.51 g, 531 mmol) wereadded to 700 ml of anhydrous THF. To the mixture were added 109 (67.32g, 177 mmol) and (n-Bu)₃SnH (114.3 ml, 425 mmol), and the mixture washeated to reflux at 85° C. for 2 hours. The reaction solution wasconcentrated under reduced pressure. To the concentrate was added methylether. The precipitated insoluble materials were removed by filtrationand the pH of the mother liquor was adjusted to 1 by adding 1N HCl. Theaqueous layer was taken and the pH was adjusted to about 9 by addingsaturated NaHCO₃, followed by extraction with ethyl acetate. The ethylacetate layers were combined, washed with a saturated saline solution,and dried over MgSO₄. MgSO₄ was removed by filtration and the filtratewas concentrated under reduced pressure. When the resulting residue waspurified by silica gel column chromatography, 39.26 g of 110 (yield 95%)was obtained from the fraction eluted with toluene/ethyl acetate (1/1).m.p. 79-80° C.

Compound 110: NMR (CDCl₃) δ: 8.66 (2H, d, J=6.2 Hz), 8.12 (1H, s), 7.85(1H, s), 7.50 (2H, dd, J=6.4, 1.8 Hz), 4.41 (2H, q, J=7.4 Hz), 1.42 (3H,t, J=7.2 Hz).

IR (CHCl₃) ν cm⁻¹: 2970, 1700, 1600, 1430, 1418, 1280, 1250, 1080.

(7) To a suspension of 110 (2.33 g, 10 mmol) in 50 ml of methanol wasadded 1N NaOH (20 ml, 20 mmol) at room temperature, and the mixture wasstirred at 50° C. for 25 minutes. After adding 20 ml of 1N HCl under icecooling, methanol was distilled away under reduced pressure. Theprecipitated solids were collected by filtration, washed in turn withisopropanol and ethyl ether, and dried to obtain 2.02 g of 111 (yield98%). m.p. ≧300° C.

Compound 111: NMR (d₆-DMSO) δ: 8.61 (2H, d, J=6 Hz), 8.51 (1H, s), 8.27(1H, s), 7.77 (2H, d, J=6 Hz).

IR (Nujol) ν cm⁻¹: 3336, 3080, 1699, 1611, 1296, 1211, 1065, 1026.

(8) H₂NCN (246 mg, 5.84 mmol) was dissolved in 15 ml of anhydrous DMF.After addition of 60% NaH (214 mg, 5.36 mmol), the mixture was stirredat room temperature for 20 minutes to prepare a solution ofNa⁺[NH—CN]⁻/DMF. Compound 111 (1.0 g, 4.87 mmol) was added to 20 ml ofanhydrous DMF, and carbonyldiimidazole (1.03 g, 5.36 mmol) was addedthereto at room temperature, and the mixture was stirred for 95 minutes.To the mixture was added dropwise the previously preparedNa⁺[NH—CN]⁻/DMF solution under ice cooling, and the mixture was stirredfor 1 hour under ice cooling. Then, 14.6 ml of hydrochloric acid wasadded and DMF was distilled away under reduced pressure. To the residuewas added 30 ml of H₂O. The precipitated solids were collected byfiltration, washed in turn with isopropanol and ethyl ether, and driedto obtain 921% of a white powder 112 (yield 82 mg). m.p. 249-251° C.

Compound 112: NMR (d₆-DMSO) δ: 8.82 (2H, d, J=6.0 Hz), 8.63 (1H, s),8.28 (2H, d, J=6.0 Hz), 8.21 (1H, s).

IR (KBr) ν cm⁻¹: 3044, 2152, 1638, 1594, 1539, 1436, 1332, 1241, 1210,816.

(1) Compound 110 (19.26 g, 82.56 mmol) was added to 500 ml of anhydroustoluene and the solution was cooled to −70° C. Under N₂ gas flow, asolution 165.12 ml (248 mmol, 1.5 M) of DIBAH/toluene was added, and themixture was stirred at −70° C. for 35 minutes. Under ice cooling,methanol (30.1 ml, 744 mmol) and H₂O (13.4 ml, 744 moles) were added.After heating to 50° C., the insoluble materials were removed byfiltration. The mother liquor was concentrated under reduced pressure toobtain 14.11 g of the desired 113 (yield 89%). m.p. 138-139° C.

Compound 113: NMR (d₆-DMSO) δ: 8.56 (2H, dd, J=6.2, 1.8 Hz), 8.09 (1H,d, J=1.6 Hz), 7.68 (2H, dd, J=6.2, 1.6 Hz), 7.51 (1H, s), 5.59 (1H, t,J=5.7 Hz), 4.67 (2H, d, J=6.0 Hz).

IR (Nujol) ν cm⁻¹: 1600, 1415, 1320, 1144, 1020, 1001, 801.

(2) To a solution of 113 (1.913 g, 10 mmol) in 250 ml of anhydrousCH₂Cl₂ was added MnO₂ (activated) (6.955 g, 80 mmol) at room temperatureunder N₂ gas flow, and the mixture was stirred for 20 hours. MnO₂ wasremoved by filtration and CH₂Cl₂ was distilled away under reducedpressure. The resulting residue was recrystallized from CH₂Cl₂/n-hexaneto obtain 1.42 g of a white powder 114 (yield 75%). m.p. 95-96° C.

Compound 114: NMR (CDCl₃) δ: 10.00 (1H, s), 8.69 (2H, d, J=6 Hz), 8.09(1H, s), 8.05 (1H, s), 7.49 (2H, d, J=6.3 Hz).

IR (CHCl₃) ν cm⁻¹: 2962, 2820, 1676, 1601, 1418, 1174, 820.

(3) To a solution of 114 (669 mg, 3.535 mmol) in 7 ml of anhydrous THFwas added Ph₃P═CHCO₂Me (1.30 g, 3.89 mmol) at room temperature under N₂gas flow, and the mixture was heated to reflux at 77° C. for 1 hour. THFwas distilled away under reduced pressure to obtain 115.

To a solution of 115 in 18 ml of methanol was added 18 ml of aqueous 1NNaOH, and the mixture was stirred at 60° C. for 1 hour. Methanol wasthen distilled away under reduced pressure, and the resulting aqueouslayer was washed three times with 100 ml of toluene. The aqueous layerwas taken and adjusted to about pH 7 by adding 118 ml of 1N HCl underice cooling. The precipitated solids were collected by filtration,washed in turn with H₂O and ethyl ether, and dried to obtain 517 mg of awhite powder 116 (yield 63%). m.p. 297-300° C. (decomposition).

Compound 116: NMR (d₆DMSO) δ: 12.51 (1H, bs), 8.62 (2H, d, J=5 Hz), 8.37(1H, s), 8.12 (1H, s), 7.75 (1H, d, J=15.0 Hz), 7.73 (2H, d, J=6 Hz),6.31 (1H, d, J=15.8 Hz).

IR (Nujol) ν cm⁻¹: 3060, 1698, 1631, 1610, 1316, 1187.

(4) To a solution of NH₂CN (182 mg, 4.32 mmol) in 5 ml of anhydrous DMFwas added 60% NaH (156 mg, 3.89 mmol) under ice cooling under nitrogengas flow, and the mixture was stirred at room temperature for 1 hour toprepare a solution of Na⁺[NH—CN]⁻/DMF. Compound 116 (500 mg, 2.16 mmol)was suspended in 15 ml of anhydrous DMF, and carbonyldiimidazole (456mg, 2.81 mmol) was added thereto at room temperature under N₂ gas flow,and the mixture was stirred for 2 hours under ice-cooling. To thesuspension was added dropwise the previously prepared Na⁺[NH—CN]⁻/DMFsolution under ice cooling, and the mixture was stirred at roomtemperature for 70 minutes. The pH was adjusted to about 7 by adding 1Nhydrochloric acid under ice cooling, and DMF was distilled away underreduced pressure. To the resulting residue was added 100 ml of ice-coldwater. The precipitated white solids were collected by filtration,washed in turn with H₂O, isopropanol and ethyl ether, and dried toobtain 471 mg of a white powder 117 (yield 86%). m.p. 248-250° C.

Compound 117: NMR (d₆-DMSO) δ: 8.66 (2H, d, J=6.75 Hz), 8.43 (1H, s),8.15 (1H, s), 7.87 (2H, d, J=15.3 Hz), 7.78 (2H, d, J=6.0 Hz), 6.39 (1H,d, J=15.6 Hz).

IR (Nujol) ν cm⁻¹: 2154, 1612, 1503, 1326, 1183, 959, 820.

Example 1

(1) To a solution of 29 (1.37 g, 4 mmol) in DMSO (15 ml) was added asolution of V-1 (3.66 g, 4.8 mmol) in acetonitrile (15 ml), and themixture was stirred at room temperature for 2 hours. After furtheradding V-1 (0.92 g, 1.2 mmol), the mixture was stirred for additional 2hours. The reaction solution was poured into a mixed solution of ethylether-water. The insoluble materials were collected by filtration,washed with water, and ethyl ether, and dried to obtain 6.03 g of I-1(yield quantitative).

Compound I-1: NMR (CDCl₃—CD₃OD) δ: 1.56 (9H, s), 1.67 (9H, s), 3.81 (3H,s), 4.59 (2H, bs), 5.1˜5.6 (5H, m), 5.77 (2H, d, J=55 Hz), 5.98 (1H, d,J=5 Hz), 6.77 (1H, bs), 6.91 (2H, d, J=8 Hz), 7.33 (2H, d, J=8 Hz), 8.03(2H, d, J=6 Hz), 8.87 (2H, d, J=6 Hz).

(2) Compound I-1 (6.03 g, 5.5 mmol) was dissolved in 40 ml ofdichloromethane and the solution was cooled to −30° C. After addition ofa solution of aluminum chloride (6.6 g, 49 mmol) in anisole (30 ml), themixture was stirred at the same temperature for 40 minutes. The reactionsolution was poured into a mixed solution of diluted hydrochloricacid-ethanol (1:1). The aqueous solution was washed with ethyl ether andthe organic solvent in the aqueous layer was distilled away underreduced pressure. The residue was then subjected to columnchromatography and eluted with an aqueous solution of 5%acetonitrile-0.05N sodium hydrogencarbonate. The eluent was concentratedunder reduced pressure and neutralized with diluted hydrochloric acid.The insoluble materials were collected by filtration and dried to obtain0.46 g of I-1′ (yield 12.8%).

Compound I-1′: NMR (d₆-DMSO+D₂O) δ: 2.97, 3.54 (2H, ABq, J=21.0 Hz),4.13, 4.40 (2H, ABq, J=16.5 Hz), 4.67, 5.22 (2H, ABq, J=15.0 Hz), 5.10(1H, d, J=5 Hz), 5.70 (2H, d, J=56 Hz), 5.74 (1H, d, J=5 Hz), 6.61 (1H,s), 7.74 (2H, d, J=7 Hz), 7.79 (1H, s), 8.67 (2H, d, J=7 Hz).

IR (KBr) ν cm⁻¹: 2252, 2151, 1773, 1671, 1636.

The intermediate I-1 is also obtainable as a salt wherein a pyridiniumcation is neutralized with an organic anion such as I⁻, depending on theconditions of isolation and purification. In such a case, it can also beconverted into the compound I-1′ by subjecting to the deprotectionreaction in the same manner.

Example 2

(1) Compound V-2 (0.91 g, 1.2 mmol) and 34 (254 mg, 1 mmol) weredissolved in DMSO (5 ml), and the mixture was stirred at roomtemperature for 2 hours. The reaction solution was poured into ethylether to precipitate the insoluble matter. The ether layer was removedby decantation and the insoluble materials were washed again with water,and ether, and dried to obtain 0.91 g of I-2 (yield 89.8%).

Compound I-2: NMR (d₆-DMSO) δ: 1.24 (3H, t, J=7 Hz), 1.51 (9H, s), 3.55,3.70 (2H, ABq, J=10 Hz), 3.71 (3H, s), 4.17 (2H, q, J=7 Hz), 5.18˜5.31(3H, m), 5.44 (2H, bs), 5.97 (1H, d, J=5 Hz), 6.91 (2H, d, J=8 Hz), 7.36(2H, d, J=8 Hz), 8.06 (1H, s), 8.15 (2H, d, J=6 Hz), 8.40 (1H, s), 8.76(2H, d, J=6 Hz).

(2) To a solution of I-2 (0.91 g, 0.90 mmol) in a mixed solution ofdichloromethane (10 ml)-nitromethane (6 ml) was added a solution ofaluminum chloride (0.72 g, 5.4 mmol) in anisole (6 ml) at −20° C., andthe mixture was stirred at the same temperature for 1 hour. The reactionsolution was poured into a mixed solution of diluted hydrochloricacid-ethanol and washed with ethyl ether. After distilling away underreduced pressure an organic solvent from the aqueous layer, the aqueoussolution was subjected to column chromatography, and eluted with anaqueous solution of 12% acetonitrile-0.05N sodium hydrogencarbonate. Theeluent was concentrated under reduced pressure and neutralized withdiluted hydrochloric acid to obtain 223 mg of I-2′ (yield 38.7%).

Compound I-2′: NMR (d₆-DMSO-D₂O) δ: 1.21 (3H, t, J=7 Hz), 3.17, 3.57(2H, ABq, J=18.0 Hz), 5.08, 5.52 (2H, ABq, J=18.0 Hz), 5.10 (1H, d, J=5Hz), 5.74 (1H, d, J=5 Hz), 8.17 (1H, s), 8.21 (2H, d, J=7 Hz), 8.36 (1H,s), 9.13 (2H, d, J=7 Hz).

IR (KBr) ν cm⁻¹: 2177, 1772, 1636.

Example 3

(1) The reaction was carried out using V-2 (910 mg, 1.2 moles) and 44(240 mg, 0.9 mmol) as starting materials in a manner similar to thatdescribed in Example 1 to obtain 0.79 of I-3 (yield 85.4%).

(2) Compound I-3 (783 mg, 0.76 mmol) was reacted with AlCl₃ inCH₂Cl₂—CH₃NO₂ to obtain 30 mg of I-3′ (yield 5.8%).

Compound I-3′: NMR (d₆-DMSO) δ: 1.21 (3H, t, J=7 Hz), 3.37, 3.52 (2H,ABq, J=18.0 Hz), 3.82 (3H, s), 4.14 (2H, q, J=7 Hz), 5.19 (1H, d, J=5Hz), 5.17, 5.38 (2H, ABq, J=15.0 Hz), 5.88 (1H, dd, J=5 Hz, 8 Hz), 6.92(1H, s), 8.05 (1H, s), 8.15 (2H, bs), 8.28 (2H, d, J=7 Hz), 8.73 (2H, d,J=7 Hz), 9.60 (2H, d, J=8 Hz), 12.12 (1H, bs).

IR (KBr) ν cm⁻¹: 2257, 2164, 1780, 1671, 1636.

Example 4

(1) To a solution of 47 (203 mg, 0.8 mmol) in DMSO (10 ml) was added V-2(789 mg, 1.03 mmol), and the mixture was stirred for 45 minutes. Theresulting solution was added dropwise to ethyl ether (500 ml) withstirring and the mother liquor was decanted to obtain an oily insolublematter. The insoluble materials were washed with H₂O, collected byfiltration, dissolved in a mixed solution of CHCl₃ and acetonitrile, anddried over MgSO₄ to obtain 830 mg of I-4.

Compound I-4: NMR (d₆-DMSO) δ: 1.24 (3H, t, J=7.0 Hz), 1.51 (9H, s),3.53 (2H, bs), 3.73 (3H, s), 4.21 (2H, q, J=7.0 Hz), 5.22 (2H, bs), 5.22(1H, d, J=5.2 Hz), 5.43 (2H, bs), 6.00 (1H, dd, J=5.2, 8.6 Hz), 6.93(2H, d, J=8.6 Hz), 7.37 (2H, d, J=8.6 Hz), 8.08 (1H, bs), 8.15 (2H, d,J=6.6 Hz), 8.33 (1H, bs), 8.77 (2H, d, J=6.6 Hz), 9.04 (1H, bs), 9.69(1H, d, J=8.6 Hz), 11.18 (1H, bs), 12.61 (1H, bs), 12.96 (1H, bs).

IR (KBr) ν cm⁻¹: 2182, 1784, 1713, 1679, 1634.

(2) To a solution of I-4 (820 mg, 0.81 mmol) in 30 ml of CH₂Cl₂ and 30ml of CH₃NO₂ was added anisole (1.06 ml, 9.75 ml). To the solution wasadded 7.29 ml of a solution of AlCl₃ in CH₃NO₂ (1M) at −20° C., and themixture was stirred at the same temperature for 30 minutes. The reactionsolution was poured into a mixture of H₂O (100 ml), ethanol (100 ml) andsodium tartrate (3.4 g, 14.8 mmol) previously stirred under ice cooling.The resulting suspension was washed with ethyl ether. The aqueous layerwas concentrated under reduced pressure and subjected to columnchromatography (40% acetonitrile/H₂O). The eluent was concentrated underreduced pressure. The precipitated insoluble materials were filtered toobtain 111 mg of I-4′ (yield 21%).

Compound I-4′: NMR (d₆-DMSO(+D₂O)) δ: 1.20 (3H, t, J=7.0 Hz), 3.09, 3.52(2H, ABq, J=19.0 Hz), 4.12 (2H, q, J=7.0 Hz), 4.96, 5.54 (2H, ABq,J=13.8 Hz), 5.06 (1H, d, J=5.0 Hz), 5.70 (1H, d, J=5.0 Hz), 8.10 (1H,bs), 7.20 (2H, d, J=7.0 Hz), 8.26 (1H, bs), 9.20 (2H, d, J=7.0 Hz).

IR (KBr) ν cm⁻¹: 2182, 1768, 1635.

Example 5

(1) To a solution of 52 (951 mg, 3.73 mmol) in 40 ml of DMSO was addedV-2 (4.24 g, 5.59 mmol), and the mixture was stirred for 30 minutes atroom temperature. The resulting reaction solution was added dropwise to200 ml of ethyl ether with stirring. The mother liquor was decanted toobtain an oily insoluble material, which was then washed with 400 ml ofH₂O. The precipitated insoluble materials were collected by filtration,and dissolved in a mixed solution of a small amount of CHCl₃ andacetonitrile. To the solution were added 100 ml of ethyl acetate and 400ml of ethyl ether, and the precipitated insoluble materials werefiltered to obtain 3.14 g of I-5 (yield 83%).

Compound I-5: NMR (d₆-DMSO) δ: 1.23 (3H, t, J=7.0 Hz), 1.50 (9H, s),2.26 (3H, s), 3.44˜3.64 (2H, m), 3.72 (3H, s), 4.20 (2H, q, J=7.0 Hz),4.20 (2H, bs), 5.22 (2H, bs), 5.23 (1H, d, J=5.4 Hz), 5.25˜5.43 (2H, m),5.99 (1H, dd, J=5.4, 8.2 Hz), 6.92 (2H, d, J=8.6 Hz), 7.21 (1H, bs),7.36 (2H, d, J=8.6 Hz), 7.78 (1H, bs), 8.05 (2H, d, J=7.2 Hz), 8.58 (2H,d, J=7.2 Hz), 9.69 (1H, d, J=8.2 Hz), 11.56 (1H, s), 12.60 (1H, bs).

IR (KBr) ν cm⁻¹: 2244, 2146, 1786, 1714, 1634.

(2) To a solution of I-5 (3.13 g, 3.09 mmol) in 30 ml of CH₂Cl₂ and 20ml of CH₃NO₂ was added anisole (4.03 ml, 37.08 mmol). After cooling to−20° C., 4.03 ml of a solution of AlCl₃ in CH₃NO₂ (1M) was added, andthe mixture was stirred at the same temperature for 40 minutes. Thereaction solution was poured into a mixture of ethanol (100 ml) and 100ml of 0.1N HCl previously stirred under ice cooling. The resultingsuspension was washed with ethyl ether. The aqueous layer wasconcentrated under reduced pressure and subjected to columnchromatography (acetonitrile/0.05N NaHCO₃ 10%). The eluent was adjustedto about pH 7 with 1N HCl, and concentrated under reduced pressure. Tothe concentrate was added 1N HCl and the precipitated insolublematerials were filtered to obtain 515 mg of I-5′ (yield 27%).

Compound I-5′: NMR (d₆-DMSO) δ: 1.16 (3H, t, J=7.2 Hz), 2.21 (3H, s),3.00, 3.56 (2H, ABq, J=18.6 Hz), 3.09 (2H, q, J=7.2 Hz), 4.09˜4.33 (2H,m), 4.70, 5.24 (2H, ABq, J=13.8 Hz), 5.08 (1H, d, J=4.6 Hz), 5.75 (1H,dd, J=4.6, 7.8 Hz), 7.74 (2H, d, J=7.0), 7.75 (1H, bs), 7.66 (2H, d,J=7.0 ), 7.46 (1H, d, J=7.8 Hz), 11.80 (1H, bs).

IR (KBr) ν cm⁻¹: 2244, 2160, 1769, 1676, 1633.

Example 6

(1) To a suspension of 55 (477 mg, 2 mmol) in 25 ml of DMSO was addedV-2 (2.73 g, 3.60 mmol), and the mixture was stirred for 7 hours at roomtemperature. The resulting reaction solution was added dropwise to 500ml of ethyl ether with stirring and the mother liquor was decanted toobtain an oily insoluble material, which was then washed with H₂O. Theprecipitated insoluble materials were collected by filtration, anddissolved in a mixed solution of CHCl₃ and acetonitrile. The solutionwas dried over MgSO₄, and added dropwise to 100 ml of ethyl acetate withstirring. When 500 ml of ethyl acetate was added, insoluble materialswere precipitated, which were filtered off to obtain 1.31 g of I-6(yield 66%).

Compound I-6: NMR (d₆-DMSO) δ: 1.23 (3H, t, J=7.0 Hz), 1.50 (9H, s),3.52 (2H, bs), 3.73 (3H, s), 4.20 (2H, q, J=7.0 Hz), 5.23 (1H, d, J=5.0Hz), 5.24 (2H, bs), 5.39 (2H, bs), 5.99 (1H, q, J=5.0, 8.2 Hz), 6.38(1H, d, J=15.2 Hz), 6.93 (2H, d, J=8.2 Hz), 7.35 (2H, d, J=8.2 Hz), 7.36(1H, bs), 7.47 (1H, d, J=15.2 Hz), 8.21 (2H, d, J=7.0 Hz), 8.25 (1H,bs), 8.68 (2H, d, J=7.0 Hz), 9.69 (1H, d, J=8.2 Hz), 12.61 (1H, s).

IR (KBr) ν cm⁻¹: 2230, 2150, 1785, 1712, 1690, 1616.

(2) To a solution of I-6 (1.30 g, 1.30 mmol) in 20 ml of CH₂Cl₂ and 20ml of CH₃NO₂ was added anisole (1.70 ml, 14.9 mmol). After cooling to−20° C., 11.7 ml of a solution of AlCl₃ in CH₃NO₂ (1M) was added, andthe mixture was stirred at the same temperature for 30 minutes. Thereaction solution was poured into a mixture of 100 ml of ethanol and 100ml of 0.1N HCl previously stirred under ice cooling. The resultingsuspension was washed with ethyl ether. The aqueous layer wasconcentrated under reduced pressure and subjected to columnchromatography (10% acetonitrile/0.05N NaHCO₃). The eluent wasconcentrated under reduced pressure and adjusted to about pH 2 with 1NHCl. The precipitated insoluble materials were filtered, and washed withisopropanol and ethyl ether to obtain 131 mg of I-6′ (yield 16%).

Compound I-6′: NMR (d₆-DMSO) δ: 1.21 (3H, t, J=7.0 Hz), 3.33, 3.54 (2H,ABq, J=18.4 Hz), 4.14 (2H, q, J=7.0 Hz), 5.17 (1H, d, J=5.0 Hz), 5.21,5.41 (2H, ABq, J=14.6 Hz), 5.86 (1H, dd, J=5.0, 8.2 Hz), 6.33 (1H, d,J=16.0 Hz), 7.18 (1H, bs), 7.24 (1H, d, J=16.0 Hz), 8.14 (2H, bs), 8.16(1H, bs), 8.19 (2H, d, J=6.2 Hz), 8.81 (2H, d, J=6.2 Hz), 9.57 (1H, d,J=8.2 Hz), 12.33 (1H, bs).

IR (KBr) ν cm⁻¹: 2232, 2148, 1772, 1673, 1619.

Example 7

(1) To a suspension of 57 (508 mg, 150 mmol) in 10 ml of DMSO was addedN,O-bis(trimethylsilyl)acetamide (403 μl, 1.63 mmol), and the mixturewas stirred. After adding V-1 (1.24 g, 1.63 mmol), the mixture wasstirred at room temperature for 95 minutes. The resulting reactionsolution was added dropwise to an aqueous 5% NaCl solution. Theprecipitated insoluble materials were collected by filtration, anddissolved in 50 ml of acetonitrile. After adding 100 ml of ethylacetate, the solution was concentrated under reduced pressure. To theconcentrate was added 100 ml of ethyl acetate, and the precipitatedinsoluble materials were filtered off to obtain 1.45 g of I-7 (yield98%).

Compound I-7: NMR (d₆-DMSO) δ: 1.51 (9H, s), 1.64 (9H, s), 3.54 (2H,bs), 3.73 (3H, s), 5.17˜5.31 (2H, m), 5.25 (1H, d, J=4.6 Hz), 5.47 (2H,bs), 5.81 (2H, d, J=55.4 Hz), 6.00 (1H, dd, J=4.6 Hz, 8.2 Hz), 6.45 (1H,d, J=15.4 Hz), 6.91 (2H, d, J=8.4 Hz), 7.35 (2H, d, J=8.4 Hz), 7.65 (1H,bs), 8.01 (1H, d, J=15.4 Hz), 8.44 (2H, d, J=6.4 Hz), 8.56 (1H, bs),8.82 (2H, d, J=6.4 Hz), 9.89 (1H, d), 12.67 (1H, bs).

IR (KBr) ν cm⁻¹: 2234, 2150, 1770, 1715, 1615.

(2) To a solution of I-7 (473 mg, 0.49 mmol) in 8 ml of CH₂Cl₂ and 3 mlof CH₃NO₂ was added anisole (640 μl, 5.89 mmol). After cooling to −4°C., 4.41 ml of a solution of TiCl₄ in CH₂Cl₂ (1M) was added, and themixture was stirred under ice cooling for 30 minutes. The reactionsolution was poured into a mixture of 50 ml of 1N NCl and ethanol (50ml) previously stirred under ice cooling. The insoluble materials werefiltered off from the resulting suspension, and dissolved in an aqueousNaHCO₃ solution. A small amount of the insoluble materials werefiltered, and the solution was separated and purified by HP-20SS columnchromatography (2-4% acetonitrile/H₂O). The eluent was adjusted to aboutpH 3.05 with 1N HCl, and concentrated under reduced pressure. Theprecipitated insoluble materials were filtered off to obtain 137 mg ofI-7′ (yield 36%).

Compound I-7′: NMR (d₆-DMSO) δ: 3.29, 3.56 (2H, ABq, J=18.0 Hz), 5.18,5.43 (2H, ABq, J=14.6 Hz), 5.19 (1H, d, J=5.0 Hz), 5.75 (2H, d, J=54.0Hz), 5.85 (1H, dd, J=5.0, 8.2 Hz), 6.34 (1H, d, J=15.6 Hz), 7.20 (1H,bs), 7.28 (1H, d, J=15.6 Hz), 8.12 (1H, bs), 8.19 (2H, d, J=6.6 Hz),8.81 (2H, d, J=6.6 Hz), 8.81 (1H, d, J=8.2 Hz).

IR (KBr) ν cm⁻¹: 2228, 2146, 1772, 1680, 1620.

Example 8

(1) To a solution of 63 (100 mg, 0.44 mmol) in 10 ml of DMSO was addedV-2 (402 mg, 0.53 mmol), and the mixture was stirred at room temperaturefor 30 minutes. The reaction solution was added dropwise to 200 ml ofethyl ether with stirring, and the mother liquor was decanted to obtainan oily insoluble material, which was dissolved in 10 ml of CH₂Cl₂. Thesolution was again added dropwise to 200 ml of ethyl ether withstirring. The precipitated insoluble materials were filtered off toobtain 37.4 mg of I-8 (yield 86%).

Compound I-8: NMR (d₆-DMSO) δ: 1.24 (3H, t, J=7.0 Hz), 1.51 (9H, s),3.44˜3.62 (2H, m), 3.70 (2H, s), 3.74 (3H, s), 4.20 (2H, q, J=7.0 Hz),5.24 (1H, d, J=5.2 Hz), 5.25 (2H, bs), 5.36 (2H, bs), 5.99 (1H, dd,J=5.2, 8.8 Hz), 6.68 (1H, bs), 6.93 (2H, d, J=8.6 Hz), 7.37 (2H, d,J=8.6 Hz), 7.97 (1H, bs), 8.10 (2H, d, J=7.0 Hz), 8.58 (2H, d, J=7.0Hz), 9.71 (1H, d, J=8.8 Hz), 11.62 (1H, bs), 11.78 (1H, bs), 12.61 (1H,bs).

IR (KBr) ν cm⁻¹: 2250, 2156, 1784, 1715.

(2) To a solution of I-8 (370 mg, 0.38 mmol) in 6 ml of CH₂Cl₂ and 6 mlof CH₃NO₂ was added anisole (490 ml, 4.51 mmol). After adding 3.38 ml ofa solution of AlCl₃ in CH₃NO₂ (1M) at −20° C., the mixture was stirredat the same temperature for 30 minutes. The reaction solution was pouredinto a mixture of a solution of CH₃COONa (863 mg, 10.5 mmol) in H₂O (100ml), ethanol (100 ml) and ethyl ether (200 ml) previously stirred underice cooling. The insoluble materials were filtered from the aqueouslayer, and dissolved in NaHCO₃ solution. A small amount of the insolublematerials were filtered. The mother liquor was separated and purified bycolumn chromatography (10% methanol/0.05N NaHCO₃). The solution wasconcentrated under reduced pressure, adjusted to about pH 6.0 with 1NHCl, and desalted to obtain 70 mg of I-8′ (yield 28%).

Compound I-8′: NMR (d₆-DMSO) δ: 1.20 (3H, t, J=7.2 Hz), 3.05, 3.49 (2H,ABq, J=18.3 Hz), 4.11 (2H, q, J=7.2 Hz), 4.84, 5.43 (2H, ABq, J=14.0Hz), 5.06 (1H, d, J=5.0 Hz), 5.67 (1H, dd, J=5.0, 8.4 Hz), 6.50 (1H,bs), 7.78 (1H, bs), 8.05 (2H, d, J=7.0 Hz), 9.01 (2H, d, J=7.0 Hz), 9.47(1H, d, J=8.4 Hz), 9.87 (1H, s), 11.54 (1H, bs).

IR (KBr) ν cm⁻¹: 2156, 1764, 1681, 1634.

In the following Examples,

represents

Example 9

(1) To a solution of 68 (234 mg, 1 mmol) in anhydrous DMSO (7 ml) wasadded V-1 (830 mg, 1.09 mmol) at room temperature under N₂ gas flow, andthe mixture was stirred for 1 hour. The reaction solution was pouredinto 70 ml of a 5% saline solution. The precipitated solids werecollected by filtration, and dissolved in a mixed solution ofacetonitrile (40 ml)/CHCl₃ (20 ml). The solution was dried over MgSO₄and concentrated under reduced pressure. The resulting residue was addedto ethyl acetate (50 ml). The resultant precipitates were collected byfiltration and dried to obtain 739 mg of a yellow powder I-9 (yield87%).

Compound I-9: NMR (d₆-DMSO) δ: 12.67 (1H, bs), 12.12 (1H, bs), 9.90 (1H,d, J=8.7 Hz), 8.58 (2H, d, J=7.2 Hz), 8.19 (2H, d, 6.9 Hz), 7.92 (1H,bs), 7.35 (2H, d, J=8.4 Hz), 7.22 (1H, s), 6.92 (2H, d, J=8.4 Hz), 5.99(1H, dd, J=11.5 Hz, 7.3 Hz), 5.90 (1H, s), 5.72 (1H, s), 5.21-5.34 (5H,m), 3.73 (3H, s), 3.43, 3.58 (2H, ABq, J=19 Hz), 1.51 (9H, s).

IR (KBr) ν cm⁻¹: 2970, 2148, 1784, 1711, 1635, 1548.

(2) A suspension of I-9 (729 mg, 0.86 mmol) in 20 ml of anhydrous CH₂Cl₂and 50 ml of anhydrous CH₃NO₂ was cooled to −3° C. In a N₂ gas flow, tothe suspension were added dropwise anisole (1.1 ml, 10.32 mmol), andthen a solution (5.16 ml, 5.16 mmol) of TiCl₄ (1 mole)/CH₂Cl₂ over 10minutes, and the mixture was stirred at 0° C. for 1 hour. The reactionsolution was then added dropwise to a mixed solution of 10 ml of 1N HCland 10 ml of a 5% saline solution under ice cooling. To the mixture wasadded ethyl ether (80 ml). The precipitated solids were collected byfiltration, washed with 1N HCl and H₂O, dissolved in NaHCO₃, andpurified by column. The portion eluted with 7% acetonitrile/0.05 NNaHCO₃ was adjusted to pH 2.9 and concentrated under reduced pressure.The precipitated solid was collected by filtration, washed in turn withH₂O, isopropanol and ethyl ether to obtain 266 mg of a pale yellowpowder I-9″ (yield 49.4%).

The same results were obtained by using a solution of AlCl₃ (1mole)/CH₃NO₂ as Lewis acid.

Compound I-9″: NMR (d₆-DMSO) δ: 12.11 (1H, bs), 8.67 (2H, d, J=6.9 Hz),8.22 (2H, d, J=7.2 Hz), 8.20 (2H, s), 7.90 (1H, s), 7.21 (1H, s), 5.90(1H, dd, J=8.4, 4.8 Hz), 5.85 (1H, s), 5.66 (1H, s), 5.38, 5.22 (2H,ABq, J=18 Hz), 5.21 (1H, d, J=5.1 Hz), 3.56, 3.36 (2H, ABq, J=18 Hz).

IR (KBr) ν cm⁻¹: 3412, 2256, 2156, 1777, 1677, 1636, 1569, 1219, 1149.

(3) In a manner similar to that described above, 7.35 g of a yellowishbrown powder I-9′ (yield 76%) was obtained from V-1(4.118 g, 9.92 mmol)and 69 (2.811 g, 9 mmol).

Compound I-9′ 3: NMR (d₆-DMSO) δ: 12.68 (1H, bs), 9.95 (1H, d, J=10.3Hz), 8.76 (2H, d, J=7.90 Hz), 8.43 (1H, s), 8.37 (2H, d, J=7.50 Hz),7.37 (3H, d, J=7.90 Hz), 6.91 (2H, d, J=8.69 Hz), 6.00 (1H, dd, J=8.77,6.32 Hz), 5.90 (1H, bs), 5.71 (1H, bs), 5.15˜5.50 (5H, m), 3.72 (3H, s),3.49, 3.59 (2H, ABq, J=19.7 Hz), 1.56 (9H, s), 1.51 (9H, s).

IR (KBr) ν cm⁻¹: 2970, 2150, 1788, 1714, 1634, 1246, 1149.

Example 10

(1) In a manner similar to that described above, 4.75 g of a yellowpowder I-10 was obtained from 70 (1.24 g, 5.0 mmol) and V-1 (4.14 g, 5.0mmol). Further, in a manner similar to that described above, 1.3 g of apale yellow powder I-10″ (yield 41%) was obtained from 1-10 (4.75 g, 5mmol).

Compound I-10: NMR (d₆-DMSO) δ: 12.67 (1H, bs), 11.84 (1H, bs), 9.90(1H, d, J=8.1 Hz), 8.60 (2H, d, J=4.9 Hz), 8.09 (2H, d, J=6.9 Hz), 7.69(1H, d, J=3.3 Hz), 7.36 (2H, d, J=8.4 Hz), 6.91 (2H, d, J=8.7 Hz), 6.04(1H, dd, J=8.4, 4.8 Hz), 5.90 (1H, bs), 5.72 (1H, bs), 5.20˜5.44 (5H,m), 3.72 (3H, s), 2.63 (3H, s), 1.51 (9H, s).

IR (KBr) ν cm⁻¹: 2970, 2150, 1784, 1711, 1633.

Compound I-10″: NMR (d₆-DMSO) δ: 11.83 (1H, bs), 9.80 (1H, d, J=8.2 Hz),8.70 (2H, d, J=6.60 Hz), 8.20 (2H, s), 8.13 (2H, d, J=6.60 Hz), 7.68(1H, d, J=3.40 Hz), 5.89 (2H, m), 5.62 (1H, s), 5.43, 5.23 (2H, ABq,J=14 Hz), 5.20 (1H, d, J=5.0 Hz), 5.59, 5.37 (2H, ABq, J=18.5 Hz), 2.64(3H, s).

IR (Nujol) ν cm⁻¹: 3184, 2144, 1773, 1675, 1633, 1215.

(2) In a manner similar to that described above, 45.1 mg of a yellowishbrown powder I-10′ was obtained from 70 (150 mg, 0.43 mmol) protectedwith Boc and V-1 (328 mg, 0.43 mmol).

Compound I-10′: NMR (CDCl₃) δ: 9.04 (1H, d, J=7.0 Hz), 8.05 (2H, d,J=6.4 Hz), 7.91 (1H, s), 7.34 (2H, d, J=10.5 Hz), 6.90 (2H, d, J=8.6Hz), 6.0 (1H, d, J=5.0 Hz), 5.93 (1H, m), 5.64 (1H, m), 5.20˜5.30 (5H,m), 3.81 (3H, s), 3.42, 3.59 (2H, ABq, J=19 Hz), 2.36 (3H, s), 1.61 (9H,s), 1.57 (9H, s).

IR (CHCl₃) ν cm⁻¹: 2980, 2250, 2154, 1769, 1716, 1634, 1543, 1245, 1149.

Example 11

(1) Compound 71 (500 mg, 2.36 mmol) was added to 20 ml of anhydrousDMSO, and V-2 (2.324 g, 3.06 mmol) was added thereto at room temperatureunder N₂ gas flow, and the mixture was stirred for 2 hours. The reactionsolution was added dropwise to 500 ml of ethyl ether to obtain aprecipitated oily product, which was dissolved in 8 ml of CH₂Cl₂. TheCH₂Cl₂ solution was slowly added dropwise to 500 ml of stirred ethylether. The resultant precipitates were filtered off and dried to obtain2.5 g of a yellow powder I-11.

Compound I-11: NMR (d₆-DMSO) δ: 12.65 (1H, bs), 12.60 (1H, bs), 9.70(1H, d, J=8 Hz), 8.68 (2H, d, J=6 Hz), 8.18 (2H, d, J=7.5 Hz), 8.15 (1H,bs), 7.45 (1H, bs), 7.37 (2H, d, J=8.5 Hz), 6.84 (2H, d, J=8.5 Hz), 5.98(1H, dd, J=10, 5 Hz), 5.12˜5.43 (5H, m), 4.20 (2H, q, J=7.25 Hz), 3.73(3H, s), 3.4˜3.7 (2H, m), 1.50 (9H, s), 1.23 (3H, t, J=7.5 Hz).

IR (CHCl₃) ν cm⁻¹: 2986, 2250, 2150, 1772, 1715, 1633.

(2) In a manner similar to that described above, 68 mg of a pale yellowpowder I-11′ (yield 37%) was obtained from I-11 (290 mg, 0.3 mmol).

Compound I-11′: NMR (d₆-DMSO) δ: 12.11 (1H, bs), 9.60 (1H, d, J=8.1 Hz),8.67 (2H, d, J=6.9 Hz), 8.23 (2H, d, J=6.9 Hz), 8.14 (2H, s), 7.90 (1H,s), 7.21 (1H, s), 5.90 (1H, dd, J=9, 5.2 Hz), 5.38, 5.21 (2H, ABq,J=15.8 Hz), 5.19 (1H, d, J=5.1 Hz), 4.14 (2H, q, J=6.9 Hz), 3.52, 3.38(2H, ABq, J=19.5 Hz), 1.21 (3H, t, J=7.05 Hz).

IR (KBr) ν cm⁻¹: 2989, 2257, 2155, 1775, 1674, 1636, 1569, 1335, 1162.

Example 12

(1) In a manner similar to that described above, 937 mg of a yellowpowder I-12 (yield 95%) was obtained from V-3 (777 mg, 1 mmol) and 71(212 mg, 1 mmol).

Compound I-12: NMR (d₆-DMSO) δ: 12.86 (1H, m), 12.64 (1H, m), 9.76 (1H,d, J=7.5 Hz), 8.70 (2H, d, J=7.5 Hz), 8.21 (1H, m), 8.19 (2H, d, J=6Hz), 7.56 (1H, s), 7.38 (2H, d, J=9 Hz), 6.92 (2H, d, J=9 Hz), 6.00 (1H,dd, J=9, 5 Hz), 5.10˜5.50 (5H, m), 4.78 (1H, m), 4.54 (2H, m), 4.32 (1H,m), 3.72 (3H, s), 1.50 (9H, s).

IR (CHCl₃) ν cm⁻¹: 3012, 2400, 2250, 1771, 1715, 1686, 1549, 1246, 1151.

(2) In a manner similar to that described above, 117 mg of a pale yellowpowder I-12′ (yield 19%) was obtained from the raw material I-12 (930mg, 0.94 mmol).

Compound I-12′: NMR (d₆-DMSO) δ: 12.11 (1H, bs), 9.67 (1H, d, J=9 Hz),8.68 (2H, d, J=6.6 Hz), 8.23 (2H, d, J=6.8 Hz), 8.16 (2H, s), 7.90 (1H,s), 7.21 (1H, s), 5.90 (1H, dd, J=9, 5 Hz), 5.38, 5.20 (2H, ABq, J=14.5Hz), 5.19 (1H, d, J=5.0 Hz), 4.75 (1H, m), 4.51 (1H, m), 4.42 (1H, m),4.27 (1H, m), 3.55, 3.26 (2H, ABq, J=19.5 Hz).

IR (KBr) ν cm⁻¹: 2256, 2154, 1776, 1676, 1636, 1569, 1334, 1161.

Example 13

(1) In a manner similar to that described above, 1.57 g of a yellowpowder I-13 was obtained from V-2 (910 mg, 1.2 mmol) and 72 (226 mg, 1mmol).

Compound I-13: NMR (d₆-DMSO) δ: 12.61 (1H, m), 11.96 (1H, m), 9.71 (1H,d, J=9 Hz), 8.60 (2H, d, J=6.5 Hz), 8.20 (1H, m), 8.11 (2H, d, J=7 Hz),7.78 (1H, s), 7.38 (2H, d, J=9.0 Hz), 6.96 (2H, d, J=9.0 Hz), 5.99 (1H,m), 5.10˜5.56 (5H, m), 4.20 (2H, q, J=6.0 Hz), 3.73 (3H, s), 2.55 (3H,s), 3.40˜3.62 (2H, m), 1.51 (9H, s), 1.23 (3H, t, J=7 Hz).

IR (CHCl₃) ν cm⁻¹: 2988, 2154, 1772, 1715, 1540, 1245, 1220.

(2) In a manner similar to that described above, 21 mg of a pale yellowpowder I-13′ (yield 3.3%) was obtained from I-13 (985 mg, 1 mmol).

Compound I-13′: NMR (d₆-DMSO) δ: 11.83 (1H, bs), 9.59 (1H, d, J=9 Hz),8.71 (2H, d, J=6.5 Hz), 8.14 (4H, m), 7.69 (1H, d, J=3.0 Hz), 5.88 (1H,dd, J=8.2, 5.1 Hz), 5.43, 5.19 (2H, ABq, J=13.6 Hz), 5.18 (1H, d, J=5.0Hz), 4.15 (2H, q, J=6.3 Hz), 3.57, 3.29 (2H, ABq, J=19 Hz), 2.64 (3H,s), 1.21 (3H, t, J=6.9 Hz).

IR (KBr) ν cm⁻¹: 2253, 2155, 1779, 1634, 1563, 1391, 1156.

Example 14

(1) In a manner similar to that described above, 2.27 g of a yellowpowder I-14 was obtained from V-2 (1.89 g, 2.48 mmol) and 67 (506 mg,2.07 mmol).

Compound I-14: NMR (d₆-DMSO) δ: 12.61 (1H, s), 12.27 (1H, s), 9.71 (1H,d, J=9 Hz), 8.78 (2H, d, J=6.9 Hz), 8.05 (2H, d, J=6.9 Hz), 7.83 (1H,s), 7.79 (1H, s), 7.71 (1H, d, J=16.5 Hz), 7.36 (2H, d, J=9 Hz), 6.92(2H, d, J=8.4 Hz), 6.38 (1H, d, J=15 Hz), 5.99 (1H, dd, J=9.8, 5 Hz),5.43 (2H, ABq, J=15.8 Hz), 5.24 (3H, d, J=5.1 Hz), 4.20 (2H, q, J=7.2Hz), 3.73 (3H, s), 3.38, 3.58 (2H, ABq, J=9.8 Hz), 1.50 (9H, s), 1.24(3H, t, J=7.05 Hz).

IR (CHCl₃) ν cm⁻¹: 2988, 2362, 1774, 1716, 1601, 1540, 1243.

(2) In a manner similar to that described above, 536 mg of a pale yellowpowder I-14′ (yield 40%) was obtained from the raw material I-14 (2.256g, 2.07 mmol).

Compound I-14′: NMR (d₆-DMSO) δ: 12.06 (1H, bs), 9.60 (1H, d, J=9 Hz),8.89 (2H, d, J=6.6 Hz), 8.03 (2H, d, J=6.9 Hz), 7.71 (1H, s), 7.57 (1H,d, J=15.3 Hz), 7.48 (1H, s), 6.25 (1H, d, J=15.6 Hz), 5.83 (1H, dd,J=8.3, 6 Hz), 5.44, 5.23 (2H, ABq, J=15 Hz), 5.18 (1H, d, J=6 Hz), 4.14(2H, q, J=6.9 Hz), 3.60, 3.37 (2H, ABq, J=18 Hz), 1.22 (3H, d, J=7.05Hz).

IR (KBr) ν cm⁻¹: 2980, 2242, 2156, 1774, 1671, 1634, 1539, 1391, 1169.

Example 15

(1) In a manner similar to that described above, 1.12 g of a yellowishbrown powder I-15 was obtained from V-2 (911 mg, 1.2 mmol) and 73 (273mg, 1 mmol).

Compound I-15: NMR (d₆-DMSO) δ: 12.60 (1H, m), 12.31 (1H, m), 9.70 (1H,d, J=9 Hz), 8.66 (2H, d, J=7.0 Hz), 8.29 (1H, s), 8.22 (2H, d, J=8.5Hz), 7.79 (1H, s), 7.61 (1H, s), 7.38 (2H, d, J=9 Hz), 6.95 (2H, d,J=9.1 Hz), 5.99 (1H, dd, J=9.5 Hz), 5.40 (2H, m), 5.24 (3H, m), 4.20(2H, q, J=7.5 Hz), 3.73 (3H, s), 3.30˜3.60 (2H, m), 1.50 (9H, s), 1.23(3H, t, J=7.5 Hz).

IR (Nujol) ν cm⁻¹: 2978, 2158, 1772, 1716, 1542, 1369, 1245.

(2) In a manner similar to that described above, 282 mg of a yellowpowder I-15′ (yield 41%) was obtained from I-15 (1.1 g, 1.0 mmol).

Compound I-15′: NMR (d₆-DMSO) δ: 12.22 (1H, bs), 9.60 (1H, d, J=9.0 Hz),8.79 (2H, d, J=6.8 Hz), 8.14 (3H, s), 7.74 (1H, s), 7.55 (1H, s), 5.87(1H, dd, J=8.5, 5 Hz), 5.41, 5.23 (2H, ABq, J=15 Hz), 5.18 (1H, d, J=5.0Hz), 4.14 (2H, q, J=7.0 Hz), 3.36˜3.60 (2H, m), 1.21 (3H, t, J=7.1 Hz).

IR (KBr) ν cm⁻¹: 2984, 2257, 2157, 1775, 1671, 1623, 1564, 1348, 1155.

Example 16

(1 ) In a manner similar to that described above, 2.26 g of a yellowpowder I-16 was obtained from V-2 (1.82 g, 2.4 mmol) and 74 (5.0 mg, 2mmol).

Compound I-16: NMR (d₆-DMSO) δ: 12.61 (1H, s), 12.30 (1H, s), 9.70 (1H,d, J=9 Hz), 8.68 (2H, d, J=6.6 Hz), 8.29 (2H, d, J=6.9 Hz), 8.13 (1H,s), 7.37 (2H, d, J=8.7 Hz), 7.31 (1H, s), 6.92 (2H, d, J=8.4 Hz), 5.99(1H, dd, J=9.5 Hz), 5.39 (2H, brs), 5.24 (3H, m), 4.20 (2H, q, J=7.2Hz), 3.73 (3H, s), 3.51 (2H, ABq, J=19.5 Hz), 2.15 (3H, s), 1.50 (9H,s), 1.23 (3H, t, J=7.2 Hz).

IR (CHCl₃) ν cm⁻¹: 2990, 2234, 2144, 1771, 1716, 1682, 1629, 1245, 1543.

(2) In a manner similar to that described above, 476 mg of a yellowpowder I-16′ (yield 36%) was obtained from I-16 (2.02 g, 2 mmol).

Compound I-16′: NMR (d₆-DMSO) δ: 12.10 (1H, bs), 9.62 (1H, d, J=9 Hz),8.73 (2H, d, J=6.9 Hz), 8.24 (2H, d, J=7.2 Hz), 8.12 (2H, s), 8.08 (1H,d, J=1.2 Hz), 7.36 (1H, s), 7.11 (1H, s), 5.86 (1H, dd, J=9.8, 5.1 Hz),5.41, 5.20 (2H, ABq, J=14 Hz), 5.18 (1H, d, J=4.8 Hz), 4.15 (2H, q,J=6.9 Hz), 3.58, 3.33 (2H, ABq, J=18 Hz), 2.09 (3H, s), 1.22 (3H, t,J=7.2 Hz).

IR (KBr) ν cm⁻¹: 2987, 2243, 2150, 1775, 1672, 1631, 1530, 1355, 1152.

Example 17

(1) To a suspension of 112 (229 mg, 1 mmol) in 7 ml of anhydrous DMSOwas added and V-1 (957 mg, 1.2 mmol) was added at room temperature underN₂ gas flow, and the mixture was stirred at room temperature for 100minutes. The insoluble materials were removed by filtration and thefiltrate was added dropwise to a 5% saline solution. The precipitatedsolids were collected by filtration, washed with a 5% saline solution,and dissolved in acetonitrile/CHCl₃ (3/1). The organic layer was takenand dried over MgSO₄. After MgSO₄ was removed by filtration, thefiltrate was concentrated under reduced pressure. The residue was thenpoured into 100 ml of ethyl acetate. The precipitated solids werecollected by filtration, and dried to obtain 344 mg of a yellow powderI-17 (yield 40%).

Compound I-17: NMR (d₆-DMSO) δ: 12.69 (1H, bs), 9.90 (1H, d, J=8.2 Hz),8.85 (2H, d, J=7 Hz), 8.71 (1H, s), 8.49 (2H, d, J=7 Hz), 8.24 (3H, s),7.37 (2H, d, J=8 Hz), 6.90 (2H, d, J=8 Hz), 5.99 (1H, dd, J=8.2, 4.8Hz), 5.94 (1H, s), 5.67 (1H, bs), 5.47 (2H, bs), 5.14-5.26 (3H, m), 3.72(3H, s), 3.40-3.70 (2H, m), 1.51 (9H, s).

IR (KBr) ν cm⁻¹: 2970, 2166, 1787, 1712, 1632, 1539, 1245, 1151, 855.

(2) In a manner similar to that described above, 25 mg of a yellowishwhite powder I-17′ (yield 10%) was obtained from I-17 (335 mg, 0.388mmol).

Compound I-17′: NMR (d₆-DMSO) δ: 9.79 (1H, d, J=8.1 Hz), 8.98 (2H, d,J=6.6 Hz), 8.66 (1H, d, J=1.8 Hz), 8.53 (2H, d, J=6.9 Hz), 8.19 (3H, s),5.89 (1H, dd, J=8.0, 5.3 Hz), 5.84 (1H, s), 5.66 (1H, s), 5.51, 5.32(2H, ABq, J=14.6 Hz), 5.20 (1H, d, J=5.1 Hz), 3.56, 3.37 (2H, ABq,J=18.3 Hz).

IR (KBr) ν cm⁻¹: 2170, 1777, 1675, 1633, 1538, 1349, 1154, 1062, 989.

Example 18

(1) In a manner similar to that described above, 1.462 g of a yellowpowder I-18 (yield 96%) was obtained from V-2 (1.749 g, 1.844 mmol) and103 (413 mg, 1.756 mmol).

Compound I-18: NMR (d₆-DMSO) δ: 14.11 (1H, bs), 12.60 (1H, bs), 9.70(1H, d, J=8.4 Hz), 8.85 (2H, d, J=6.8 Hz), 8.47 (2H, d, J=6.8 Hz), 7.49(1H, s), 7.35 (2H, d, J=8.6 Hz), 6.91 (2H, d, J=8.6 Hz), 5.99 (1H, dd,J=8.6, 5.0 Hz), 5.43, 5.51 (2H, ABq, J=16.5 Hz), 5.16-5.33 (3H, m), 4.19(2H, q, J=7.0 Hz), 3.72 (3H, s), 3.60, 3.47 (2H, ABq, J=19 Hz), 1.50(9H, s), 1.23 (3H, t, J=7.2 Hz).

IR (KBr) ν cm⁻¹: 2970, 2154, 1789, 1713, 1636, 1538, 1341, 1245, 1151,1034.

(2) In a manner similar to that described above, 330 mg of a whitepowder I-18′ (yield 32%) was obtained from I-18 (1.452 g, 1.677 mmol).

Compound I-18: NMR (d₆-DMSO) δ: 14.09 (1H, bs), 9.60 (1H, d, J=8.4 Hz),8.92 (2H, d, J=6.9 Hz), 8.51 (2H, d, J=6.9 Hz), 8.12 (2H, s), 7.47 (1H,s), 5.91 (1H, dd, J=8.4, 4.8 Hz), 5.52, 5.38 (2H, ABq, J=14.9 Hz), 5.19(1H, d, J=5.1 Hz), 4.14 (2H, q, J=7.2 Hz), 3.56, 3.41 (2H, ABq, J=19.5Hz), 1.21 (3H, t, J=7.05 Hz).

IR (KBr) ν cm⁻¹: 2970, 2158, 1774, 1671, 1636, 1526, 1403, 1345, 1154,1036.

Example 19

(1) To a suspension of 117 (306 mg, 1.2 mmol) in 6 ml of anhydrous DMSOwas added dropwise N,O-bis(trimethylsilyl)-acetamide (292 μl, 1.2 mmol)under N₂ gas flow with stirring. After adding V-2 (1.195 g, 1.26 mmol),the mixture was stirred at room temperature for 85 minutes. The reactionsolution was then added dropwise to 65 ml of a 5% saturated salinesolution. The precipitated solids were collected by filtration andwashed with H₂O. The yellow solids were dissolved in acetonitrile/CHCl₃(80 ml/30 ml) and the solution was dried over MgSO₄. After MgSO₄ wasremoved by filtration, the filtrate was concentrated under reducedpressure. The residue was poured into 50 ml of ethyl acetate, and 100 mlof ethyl ether was further added. The precipitated solids were collectedby filtration, and dried to obtain 840 mg of a yellow powder I-19 (yield79%).

Compound I-19: NMR (d₆-DMSO) δ: 12.60 (1H, bs), 9.69 (1H, d, J=8.4 Hz),8.91 (2H, d, J=7.2 Hz), 8.78 (1H, s), 8.44 (2H, d, J=6.9 Hz), 8.19 (1H,s), 7.63 (1H, d, J=15.6 Hz), 7.35 (2H, d, J=8.7 Hz), 6.91 (2H, d, J=8.7Hz), 6.39 (1H, d, J=15.6 Hz), 5.99 (1H, dd, J=8.6 Hz, 5.0 Hz), 5.53,5.46 (2H, ABq, J=13.5 Hz), 5.17-5.26 (3H, m), 4.20 (2H, q, J=7.2 Hz),3.72 (3H, s), 3.54, 3.59 (2H, ABq, J=17.3 Hz), 1.50 (9H, s), 1.23 (3H,t, J=7 Hz).

IR (KBr) ν cm⁻¹: 2970, 2228, 2148, 1788, 1713, 1630, 1537, 1514, 1369,1245, 1153, 1033.

(2) In a manner similar to that described above, 232 mg of a yellowishwhite powder I-19′ (yield 37%) was obtained from I-19 (831 mg, 0.94mmol).

Compound I-19′: NMR (d₆-DMSO, D₂O) δ: 9.62 (1H, d, J=7.5 Hz), 9.00 (2H,d, J=6.0 Hz), 8.70 (1H, s), 8.44 (2H, d, J=4.8 Hz), 8.12 (1H, s), 7.58(1H, d, J=15.3 Hz), 6.35 (1H, d, J=15.2 Hz), 5.87 (1H, d, J=6 Hz), 5.54,5.33 (2H, ABq, J=15 Hz), 5.17 (1H, d, J=6 Hz), 4.13 (2H, q, J=7.5 Hz),3.77, 3.37 (2H, ABq, J=18 Hz), 1.22 (3H, t, J=6.8 Hz).

IR (KBr) ν cm⁻¹: 2257, 2158, 1776, 1674, 1623, 1536, 1357, 1156, 1038.

Example 20

(1) In a manner similar to that described above, 393 mg of a yellowpowder I-20 (quantitative) was obtained from 96 (E) (123 mg, 0.488 mmol)and V-2 (458 mg, 0.58 mmol).

Compound I-20: NMR (d₆-DMSO) δ: 12.60 (1H, bs), 12.49 (1H, bs), 9.69(1H, d, J=8.1 Hz), 8.68 (2H, d, J=6.9 Hz), 8.24 (2H, d, J=6.6 Hz), 8.22(1H, s), 7.41 (1H, s), 7.35 (2H, d, J=8.7 Hz), 6.92 (2H, d, J=8.7 Hz),6.27 (1H, d, J=0.6 Hz), 5.99 (1H, dd, J=8.25, 4.95 Hz), 5.41, 5.38 (2H,ABq, J=9 Hz), 5.16-5.25 (3H, m), 4.20 (2H, q, J=7.2 Hz), 3.73 (3H, s),3.57, 3.49 (2H, ABq, J=17.3 Hz), 1.50 (9H, s), 1.24 (3H, t, J=7.05 Hz).

IR (KBr) ν cm⁻¹: 2972, 2230, 2144, 1786, 1711, 1633, 1610, 1543, 1245,1153, 1033, 931.

(2) In a manner similar to that described above, 75 mg of a yellowishgreen powder I-20′ (yield 26%) was obtained from I-2 (382 mg, 0.43mmol).

Compound I-20′: NMR (d₆-DMSO) δ: 12.26 (1H, bs), 9.55 (1H, d, J=7.8 Hz),8.84 (2H, d, J=6 Hz), 8.21 (2H, d, J=6 Hz), 8.13 (2H, s), 8.08 (1H, s),7.25 (1H, s), 6.31 (1H, s), 5.84 (1H, dd, J=9, 5.4 Hz), 5.39, 5.16 (2H,ABq, J=14 Hz), 5.14 (1H, d, J=5.4 Hz), 4.14 (2H, q, J=6.3 Hz), 3.58,3.26 (2H, ABq, J=20 Hz), 2.50 (3H, s), 1.21 (3H, t, J=6.9 Hz).

IR (KBr) ν cm⁻¹: 2238, 2147, 1775, 1676, 1635, 1605, 1525, 1353, 1154,1037, 933.

Example 21

In a manner similar to that described above, 485 mg (99%) of a yellowishbrown powder I-21 was obtained from 96 (Z) (162 mg, 0.642 mmol) and V-2(552 mg, 0.706 mmol).

Compound I-21: NMR (d₆-DMSO) δ: 12.60 (1H, bs), 12.42 (1H, bs), 9.69(1H, d, J=8.1 Hz), 8.65 (2H, d, J=6.6 Hz), 8.22 (2H, dd, J=6.6, 0.6 Hz),8.16 (1H, d, J=3.0 Hz), 7.36 (2H, d, J=8.4 Hz), 7.33 (1H, d, J=3.0 Hz),6.92 (2H, d, J=8.4 Hz), 5.98 (1H, dd, J=8.9, 4.5 Hz), 5.71 (1H, s),5.39, 5.37 (2H, ABq, J=8 Hz), 5.13-5.26 (3H, m), 4.20 (2H, q, J=6.9 Hz),3.73 (3H, s), 3.48, 3.56 (2H, ABq, J=18.8 Hz), 2.24 (3H, s), 1.50 (9H,s), 1.23 (3H, t, J=7.2 Hz).

IR (KBr) ν cm⁻¹: 2970, 2240, 2144, 1787, 1712, 1632, 1546, 1514, 1246,1154, 1033, 932.

Test 1

The minimal inhibitory concentration (MIC) was determined by an agardilution method. That is, 1.0 ml each of an aqueous solution of a testcompound diluted in series was poured into a petri dish. Trypticase soyagar (9 ml) was poured into the solution, and mixed. A suspension (about10⁶ CFU/ml) of test bacteria was smeared on the mixed agar plate. Afterculturing at 37° C. overnight, the minimum concentration of the testcompound required to completely inhibit the growth of the test bacteriawas taken as MIC.

Test Bacteria:

Gram-positive bacteria; S. pyogenes C-203, S. agalactiae ATCC13813, S.pneumoniae Type I, S. pneumoniae SR16675 (PC-R), S. mitis ATCC9811Gram-negative bacteria: K. pneumoniae SR1, P. mirabilis PR-4 and P.vulgaris CN-329.

Results:

TABLE 1 MIC(μg/ml) Inoculation Compound of amount: the present invention10⁶ CFU/ml I-1′ I-9″ Control compound Test bacteria (Example 1) (Example9) (1) (2) Gram-positive bacteria S. pyogenes 0.006 0.006  0.013 0.006C-203 S. agalactiae 0.025 0.013 0.1  0.025 ATCC13813 S. pneumoniae 0.0130.006 0.05 0.006 Type I S. pneumoniae 0.2  0.2  0.78 0.39  SR16675(PC-R) S. mitis 0.05  0.025 0.1  0.025 ATCC9811 Gram-negative bacteriaK. pneumoniae 0.006 0.006 0.05 0.013 R1 P. mirabilis 0.013 0.013 0.1 0.05  PR-4 P. vulgaris 0.013 0.013 0.1  0.025 CN-329 (Control compound)(1)

(2)

As is apparent from Table 1, the compound of the present inventionexerts excellent and well-balanced antibacterial activity to typicalstrains of pathogenic bacteria, which are considered important fromclinical point of view.

Test 2

The blood half-life and the treating effect on infections in mouse wereexamined by using the compound I-9″ and control compound (1) as used inTest 1 above.

The results are shown in Table 2 and Table 3.

TABLE 2 Half-life in Blood (h) I-9″ Control compound (1) Mouse 0.96 0.39Monkey 2.48 1.28

For the exertion of antibacterial activity to Pseudomonas, it isnecessary that a drug contacts with bacteria for a long-term.Accordingly, the longer the blood half-life (T_(1/2)), the moreadvantageous. The above results show that the compound of the presentinvention is effective on infectious diseases caused by Pseudomonas.

TABLE 3 Treating Effect on Infections in Mouse, ED₅₀ (mg/kg) I-9″Control compound (1) S. aureus Smith 0.41 0.76 S. pneumoniae Type I0.042 0.69 P. vulgaris GN-329 0.009 0.61 P. aeruginosa SR24 0.38 1.52 P.aeruginosa E-2 0.79 4.73

The above results show that the compound of the present invention iseffective in vivo.

What we claim is:
 1. A cephem compound shown by the formula I:

wherein Acyl is selected from the group consisting of formyl, (C₁-C₆)alkylcarbonyl, (C₃-C₅) alkenoyl, (C₃-C₁₀) cycloalkyl-carbonyl,(C₅-C₆)cycloalkenyl-carbonyl, arylcarbonyl, aralkyl carbonyl, 5- or6-membered aromatic heterocyclic carbonyl, 5- or 6-membered aromaticheterocyclic acetyl, alkoxycarbonyl, aryloxycarbonyl,aralkyloxycarbonyl, aminoalkylcarbonyl, monoalkylaminoalkylcarbonyl, anddialkylaminoalkylcarbonyl, optionally substituted with one to threesubstituents independently selected from the group consisting of amino,nitro, halogen, hydroxy, oxo, carbamoyl, (C₁-C₄) alkyl, (C₁-C₄) alkoxy,(C₁-C₄) alkoxycarbonyl, (C₁--C₄) alkoxyimino optionally substituted withcarboxyl or halogen, hydroxyimino and4-ethyl-2,3-dioxopiperadinocarbonylamino, or acyl is represented byformula III:

 wherein X is CH or N; Y is an optionally protected amino; and Z ishydrogen or an optionally substituted hydrocarbon group; Het is a mono-or polycyclic heterocyclic group comprising one or more hetero atomsselected from the group consisting of N, O and S which may be the sameor different from each other; R¹ is hydrogen, a straight or branchedlower alkyl which is optionally substituted with 1 to 3 substituentsindependently selected from the group consisting of C₂₋₆ alkenyl,cycloalkyl, aryl which is optionally substituted with hydroxy, C₁₋₄alkyl, or C₁₋₄ alkoxy, an aromatic heterocyclic group comprising 1 to 4hetero atoms selected from the group consisting of N, O, and S, a bi- ortricyclic aromatic condensed heterocyclic group comprising 1 to 5 heteroatoms selected from the group consisting of N, O, and S, which is formedby condensing one or two 5- or 6-membered-aromatic heterocyclic groupscomprising 1 to 4 hetero atoms selected from N, O, and S, a non-aromaticheterocyclic group, amino, mono- or di-alkyl amino, amidino, C₁₋₆alkanoyl, carbamoyl, mono- or di-lower alkylcarbamoyl, sulfamoyl, mono-or di-lower alkyl sulfamoyl, carboxyl, lower alkoxycarbonyl, hydroxyl,lower alkoxy, lower alkenyloxy, cycloalkyloxy, aralkyloxy, aryloxy,mercapto, lower alkylthio, aralkylthio, arylthio, sulfo, cyano, azide,nitro, nitroso and halogen, or a straight or branched lower alkenylwhich is optionally substituted with 1 to 3 substituents independentlyselected from the group consisting of C₂₋₆ alkenyl, cycloalkyl, arylwhich is optionally substituted with hydroxy, C₁₋₄ alkyl, or C₁₋₄alkoxy, an aromatic heterocyclic group comprising 1 to 4 hetero atomsselected from the group consisting of N, O, and S, a bi- or tricyclicaromatic condensed heterocyclic group comprising 1 to 5 hetero atomsselected from the group consisting of N, O, and S, which is formed bycondensing one or two 5- or 6-membered aromatic heterocyclic groupscomprising 1 to 4 hetero atoms selected from N, O, and S, a non-aromaticheterocyclic group, amino, mono- or di-alkyl amino, amidino, C₁₋₆alkanoyl, carbamoyl, mono- or di-lower alkylcarbamoyl, sulfamoyl, mono-or di-lower alkyl sulfamoyl, carboxyl, lower alkoxycarbonyl, hydroxyl,lower alkoxy, lower alkenyloxy, cycloalkyloxy, aralkyloxy, aryloxy,mercapto, lower alkylthio, aralkylthio, arylthio, sulfo, cyano, azide,nitro, nitroso and halogen; A is a lower alkylene which is optionallysubstituted with 1 to 3 substituents independently selected from thegroup consisting of C₂₋₆ alkenyl, cycloalkyl, aryl which is optionallysubstituted with hydroxy, C₁₋₄ alkyl, or C₁₋₄ alkoxy, an aromaticheterocyclic group comprising 1 to 4 hetero atoms selected from thegroup consisting of N, O, and S, a bi- or tricyclic aromatic condensedheterocyclic group comprising 1 to 5 hetero atoms selected from thegroup consisting of N, O, and S, which is formed by condensing one ortwo 5- or 6-membered aromatic heterocyclic groups comprising 1 to 4hetero atoms selected from N, O, and S, a non-aromatic heterocyclicgroup, amino, mono- or di-alkyl amino, amidino, C₁₋₆ alkanoyl,carbamoyl, mono- or di-lower alkylcarbamoyl, sulfamoyl, mono- ordi-lower alkyl sulfamoyl, carboxyl, lower alkoxycarbonyl, hydroxyl,lower alkoxy, lower alkenyloxy, cycloalkyloxy, aralkyloxy, aryloxy,mercapto, lower alkylthio, aralkylthio, arylthio, sulfo, cyano, azide,nitro, nitroso and halogen, a lower alkenylene which is optionallysubstituted with 1 to 3 substituents independently selected from thegroup consisting of C₂₋₆ alkenyl, cycloalkyl, aryl which is optionallysubstituted with hydroxy, C₁₋₄ alkyl, or C₁₋₄ alkoxy, an aromaticheterocyclic group comprising 1 to 4 hetero atoms selected from thegroup consisting of N, O, and S, a bi- or tricyclic aromatic condensedheterocyclic group comprising 1 to 5 hetero atoms selected from thegroup consisting of N, O, and S, which is formed by condensing one ortwo 5- or 6-membered aromatic heterocyclic groups comprising 1 to 4hetero atoms selected from N, O, and S, a non-aromatic heterocyclicgroup, amino, mono- or di-alkyl amino, amidino, C₁₋₆ alkanoyl,carbamoyl, mono- or di-lower alkylcarbamoyl, sulfamoyl, mono- ordi-lower alkyl sulfamoyl, carboxyl, lower alkoxycarbonyl, hydroxyl,lower alkoxy, lower alkenyloxy, cycloalkyloxy, aralkyloxy, aryloxy,mercapto, lower alkylthio, aralkylthio, arylthio, sulfo, cyano, azide,nitro, nitroso and halogen or a single bond; B is —NH— or a single bond;and D is a single bond or a group of the formula:

 or an ester, a salt or a hydrate thereof.
 2. A pharmaceuticalcomposition comprising a compound of claim 1 in association withpharmaceutically acceptable carriers.
 3. The compound of claim 1,wherein the Acyl is represented by the formula III:

wherein X is CH or N; Y is an optionally protected amino; and Z ishydrogen or an optionally substituted hydrocarbon group.
 4. The compoundof claim 1, wherein Het is a 5- or 6-membered hetero cyclic groupcomprising one to four hetero atoms selected from the group consistingof N, O and S which may be the same or different from each other.
 5. Thecompound of claim 1, wherein Het is a group of the formula IV:


6. The compound of claim 1, wherein A is a single bond or a vinyl group;B is a single bond; and D is a single bond.
 7. The compound of claim 1,wherein Acyl is a group of the formula III:

wherein X is CH or N, Y is an optionally protected amino and Z ishydrogen or an optionally substituted hydrocarbon group; Het is a 5- or6-membered hetero cyclic group comprising one to four hetero atomsselected from the group consisting of N, O and S which may be the sameor different from each other; A is a single bond or a vinyl group; B isa single bond; and D is a single bond.